MXPA99011977A - Systems and methods for harvesting mononuclear cells by recirculation of packed red blood cells - Google Patents

Abstract

A blood separation system operates in a first mode to convey whole blood into the inlet region (48) of a blood processing chamber (38) for centrifugal separation into packed red blood cells, a plasma constituent, and an interface, which carries mononuclear cells, between the packed red blood cells and the plasma constituent. The system removes packed red blood cells and the plasma constituent from the chamber (38), while maintaining the interface within the chamber (38). The system operates in a second mode to remove the interface from the chamber (38) by conveying packed red blood cells into the inlet region (48). An outlet path (46) conveys the removed interface from the chamber (38). The outlet path (46) includes a first sensing element (OS) to locate mononuclear cells in the removed interface and provide a sensed output upon locating mononuclear cells.

Description

SYSTEMS AND METHODS TO COLLECT MONONUCLEAR CELLS THROUGH THE RECIRCULATION OF CONDENSED RED BLOOD CELLS OF THE BLOOD Field of the Invention The invention relates to systems and centrifugal processing apparatus.

Background of the Invention Currently, blood collection organizations commonly separate whole blood by centrifugation in its various therapeutic components, such as red blood cells, platelets, and plasma. Conventional blood processing systems and methods use durable centrifugal equipment in association with sterile, single-use processing chambers, usually made of plastic. The centrifugal equipment, introduces the whole blood in these cameras while they rotate, to create __ a centrifugal field. The whole blood is separated within the spin chamber under the influence of the centrifugal field in the red blood cells of high density and in platelet-enriched plasma. An intermediate layer of EF: 3? 28Q leukocytes forms the interface between red blood cells and platelet-enriched plasma. Mononuclear cells (MNC) are present in the interface.

Brief description of the invention The invention provides systems and methods for the collection of mononuclear cells from whole blood. The systems and methods operate in a first mode for transporting the whole blood in the inlet region of the blood processing chamber for centrifugal separation in the condensed red blood cells of the blood, in a plasma constituent, and an interface between the blood cells. the condensed red blood cells of the blood and the plasma constituent. The interface carries the mononuclear cells • designated for the collection. In the first mode, the systems and methods remove the "condensed red blood cells from the blood and the plasma constituent of the chamber, while maintaining the interface inside the chamber." The system and methods operate in a second mode to remove the camera interface by conducting the red blood cells condensed from the blood in the input region.The introduction of the red blood cells condensed from the blood increases the hematocrit in the input region, causing the floating of the mononuclear cells to the surface of the interface , where they are presented for collection, an exit path carries the interface removed from the camera, the output path includes a detector element, the detector element locates the mononuclear cells at the removed interface and provides a detected output for the location of the mononuclear cells In a preferred embodiment, the exit path includes a container in a Flow ection downstream of the detector element. In this embodiment, the systems and methods direct the collection of mononuclear cells in the container, based at least in part, on the detected output. Other features and advantages of the invention will be clear upon review of the following specification, drawings and appended claims.

BRIEF DESCRIPTION OF THE DRAWINGS Figure 1 is a view of the lateral section of the blood centrifuge having a separation chamber including the features of the invention; Figure 2 shows that the coil member associated with the centrifuge shown in Figure 1, with an associated processing vessel covered thereon for use,; Figure 3A is a perspective view of the centrifuge shown in Figure 1, with the coil and concave elements rotated in their access position; Figure 3B is a perspective view of the coil and concave elements in their mutual separation condition to make it possible to secure the processing container shown in Figure 2 on the coil member; Figure 4 is a plan view of the processing vessel shown in Figure 2; Figure 5 is a perspective view of a fluid circuit associated with the processing vessel, which comprises cartridges mounted in association with the pump stations in the centrifuge; Figure 6 is a schematic view of the fluid circuit shown in Figure 5; Figure 7 is a perspective view of the opposite side of a cartridge forming part of the fluid circuit shown in Figure 6; Figure 8 in a perspective view of the front side of the cartridge shown in Figure 7; Figure 9 is a schematic view of the flow channels and valve stations formed within the cartridge shown in Figure 7; Figure 10 is a schematic view of a pump station in order to receive a cartridge of the type shown in Figure 7; Figure 11 is a schematic view of the cartridge shown in Figure 9. mounted on the pump station shown in Figure 10; Figure 12 is a perspective view of a cartridge and a pump station that are part of the fluid circuit shown in Figure 6; Figure 13 is a top view of a peristaltic pump forming part of the fluid circuit shown in Figure 6, with the pump rotor in the retracted position; Figure 14 is a top view of a peristaltic pump forming part of the fluid circuit shown in Figure 6, with the rotor of the pump in an extended position that couples the pump tubing; Figure 15 is a schematic top view of the separation chamber of the centrifuge shown in Figure 1, positioned to show the radial contours of the high G and low G walls; Figures 16A and 16B show somewhat schematically a portion of the collection zone of platelet enriched plasma in the separation chamber, in which the high wall surface G forms a tapered wedge to contain and control the position of the interface between red blood cells and platelet-enriched plasma; Figure 17 is in a certain way a schematic view of the inside of the processing chamber, seen from the low wall G towards the high wall G in the region where the whole blood enters the processing chamber for separation in the red blood cells of blood and platelet-enriched plasma, and where platelet-enriched plasma is collected in the processing chamber; Figure 18 is a schematic view showing the dynamic flow conditions established to confine and "park" the MNCs within the blood separation chamber shown in Figure 17; Figure 19 is a schematic view of the process controller which configures the fluid circuit shown in Figure 6 to conduct a prescribed MNC collection procedure.

Figure 20 is a flow diagram showing the different cycles and phases of the collection procedure of the MNCs directed by the controller shown in Figure 19; Figure 21 is a schematic view showing the conduction of blood components and fluids in the circuit shown in Figure 6 during the preliminary process cycle of the procedure shown in Figure 20; Figure 22 is a schematic view showing the conduction of blood components and fluids in the circuit shown in Fig. 6 during the accumulation phase of the MNC of the procedure shown in Figure 20; Figure 23 is a schematic view showing the conduction of blood components and fluids in the circuit shown in Figure 6 during the collection phase of the PRBC of the procedure shown in Figure 20; Figure 24A is a schematic view showing the conduction of blood components and fluids in the circuit shown in Figure 6 at the beginning of the deposition phase __ of the MNCs of the procedure shown in Figure 20; Figure 24B is a schematic view showing the conduction of blood components and fluids in the circuit shown in Figure 6 during the deposition phase of the MNC of the procedure shown in Figure 20; Figure 24C is a schematic view showing the conduction of blood components and fluids in the circuit shown in Figure 6 at the end of the MNC deposition phase of the procedure shown in Figure 20; Figure 25 is a schematic view showing the conduction of blood components and fluids in the circuit shown in Figure 6 during the flow phase of the PRP of the procedure shown in Figure 20; Figure 26 is a schematic view showing the conduction of blood components and fluids in the circuit shown in Figure 6 during the suspension phase of the MNCs of the procedure shown in Figure 20; Figure 27 is a schematic view showing the conduction of blood components and fluids in the circuit shown in Figure 6 during the cleaning phase of the procedure shown in Figure 20; Figure 28 is a schematic view of the optical detector used in association with the circuit shown in Figure 6 to detect and quantify the region for MNC collection; L Figure 29 is an alternative modality of a fluid circuit suitable for the concentration and collection of the MNCs; Figure 30 is a schematic view showing the conduction of blood components and fluids in the circuit shown in Figure 29 during the collection phase of the PRBC of the procedure shown in Figure 20; and Figure 31 is a schematic view showing the conduction of the blood components and fluids in the circuit shown in Figure 29 during the deposition phase of the MNCs of the procedure shown in Figure 20. The -invention may be included in several. forms without departing from his spirit or essential characteristics. The scope of the invention is defined in the appended claims, instead of the specific description that precedes them. All modalities that fall within the meaning and range of equivalence of the claims are therefore intended to be adopted by the claims.

DESCRIPTION OF THE PREFERRED EMBODIMENTS I. The Centrifuge Figure 1 shows a blood centrifuge 10 having a blood processing chamber 12 suitable for the collection of mononuclear cells (MNC) from whole blood. The boundaries of the chamber 12 are formed by a flexible processing vessel 14 carried within an annular groove 16 between a rotating spool member 18 and the concave element 20. In the illustrated and preferred embodiment, the processing vessel 14 takes the shape of an elongated tube (see Figure 2) that is covered over the coil member 18 before use. Additional details of the centrifuge 10 are set forth in U.S. Patent 5,370,802, entitled "In aced Yield Platelet Systems and Methods" which is incorporated herein by reference. The coil and concave elements 18 and 20 rotate on a rocker 22 between a straight position, as shown in Figures 3A and 3B, and a suspended position, as shown in Figure 1. When the elements of vat and coil 18 Y are in the straight position are presented to be accessed by the user. A mechanism allows the tub and coil elements 18 and 20 to be opened, as shown in Figure 3B, so that the operator can cover the container 14 on the coil member 20, as shown in Figure 2. The pins 150 of the coil member 20 engage the cuts in the container 14 to secure the container 14 in the coil member 20. When closed, the coil and concave elements 18 and 20 can rotate in the suspended position shown in Figure 1. In In operation, the centrifuge 10 rotates the suspended coil and concave elements 18 and 20 on an axis 28, creating a centrifugal field within the processing chamber 12. Additional details of the mechanism for causing the relative movement of the coil and concave elements 18 and 20 are described in U.S. Patent 5,360,542 entitled "Centrifuge With Separable Bole and Spool Elements Providing Access to the Separation Chamber", which is incorporated herein by reference The radial limits of the centrifugal field (see Figure 1) are formed by the inner wall 24 of the concave element 18 and the outer wall 26 of the coil element 20. The concave inner wall 24 defines the high wall G. The outer wall of the coil 26 defines the low wall G.

II. Processing Vessel In the illustrated embodiment (see Figure 4), a first peripheral seal 42 forms the outer rim of the container 14. A second inner seal 44 generally extends "parallel to the rotational axis 28, dividing the container 14 into two compartments 38 and 40. In use, the whole blood is centrifugally separated in the compartment 38. In use, the compartment 40 carries a liquid, like the saline, to counterbalance the compartment 38. In the embodiment shown in Figure 4, the compartment 38 is greater than the compartment 40 by a volumetric ratio of approximately 1 to 1.2. Three ports 46, 48 ,. and 50 communicate with the processing compartment 38, to drive the whole blood and its components. Two additional ports 52 and 54 communicate with the ballast compartment 40 to drive the counterbalanced fluid.

III. Fluid Processing Circuit A fluid circuit 200 (see Figure 4) is coupled to the container 14. Figure 5 shows a general scheme of the fluid circuit 200, in terms of a flexible pipe arrangement, of a liquid source and collection containers, of in-line pumps, and _ clamps, all of which will be described in greater detail later. Figure 6 shows the details of the fluid circuit 200 in schematic form. In the illustrated embodiment, the cartridges, left, middle, and right, respectively 23L, 23M, and 23R, centralize several of the valve and pump functions of the fluid circuit 200. The left, middle, and right cartridges 23L, 23M, and 23R are accompanied with the left, middle, and right pumping stations in the centrifuge 10, which are designated, respectively, PSL, PSM, and PSR.

A. The Cartridges Each cartridge 23L, 23M, and 23R are constructed in the same way, so that only a description of a .cartucho 23L is applicable to all cartridges. The Figures 7 and 8 show the structural details of the cartridge 23L. The cartridge 23L comprises a molded plastic body 202. The liquid flow channels 208 are integrally molded on the front side 204 of the body 202. A rigid board 214 covers and seals the front side body 204. The valve stations 210 are molded on the opposite side 206 of the cartridge body 202. A flexible diaphragm 212 covers and seals the opposite side 206 of the body 202. Figure 9 shows schematically a representative arrangement of the flow channels 208 and the valve stations 210 for each cartridge. As shown, channels Cl through C6 intersect to form a star array and radiate from a central hub H. Channel C7 intercepts channel C5; channel C8 intercepts channel C6; channel C9 intercepts C3; and the CÍO channel intercepts channel C2. Of course, other channel models can be used. In this arrangement, the valve stations VSl, VS2, VS9, and VS10 are located in, respectively, channels C2, C3, C5, and C6 immediately adjacent to their common intersection in the hub H. Valve stations VS3, VS4, VS5, VS6, VS7, and VS8 are located on the outer extremities of channels C8, Cl, C2, C5, C4, and C3, respectively. Each cartridge 23L carries a loop of the upper flexible pipe UL extending outside the cartridge 23L between the channels C7 and C6 and a loop of the lower pipe LL extending out of the cartridge between the channels C3 and CIO. In use, the UL and LL tube loops couple the peristaltic pump rotors of the pumps to the associated pump station.

B. Pumping Stations The PSL, PSM, and PSR pumping stations are, like the 23L, 23M, and 23R cartridges, identically constructed, so that only one description of a PSL station is applicable to all. Figure 12 shows the structural details of the left pump station PSL. Figure 10 shows the left pumping station PSL in a more schematic form. The PSL station includes two peristaltic pumps, for a total of six pumps in circuit 200, which are designated Pl to P6 (see Figure 6). The PSL station also includes an array of ten valve actuators (which are shown in Figure 10), for a total of thirty valve actuators in the circuit 200 which are designated VAI to VA30 (see Figure 6). In use (see Figure 11), the UL and LL tube loops of the 23L cartridge couple the Pl and P2 pumps of the PSL left pump station. In a similar manner (as shown in Figure 6), UL and LL loops of medium cartridge 23M couple pumps P3 and P4. The UL and LL tube loops of the right cartridge 23L couple the P5 and P6 pumps. As shown in Figure 11, the valve stations VSl to VS10 of the cartridge 23L are aligned with the valve actuators VI to VIO of the left pump station PSL. As shown in Figure 6, the valve stations of the middle and right cartridges 23M and 23R equally align respectively with the valve actuators of the medium and right pump stations PSM and PSR. Table 1 below summarizes the operative association of the valve actuators VI to V30 of the pumping station to the valve stations of the cartridge VSl to VS10 shown in Figure 6.

Table 1: Alignment of Cartridge Valve Stations to Valve Actuators Case Cameras Half case It is left valve 23L 23M right 23R VSl Valve Valve actuator valve I actuator Vil actuator V21 VS2 Valve Valve Actuator valve V2 Actuator V12 Actuator V22 VS3 Valve Valve Actuator V3 Actuator V13 Actuator V23 VS4 Valve Valve Actuator valve V4 Actuator VI 4 Actuator V24 VS5 Valve Valve Actuator valve V5 Actuator V15 Actuator V25 VS6 Valve Valve Actuator Valve V6 Actuator VI 6 Actuator V26 Table 1: Alignment of Cartridge Valve Stations to Valve Actuators (Continued) Case Cameras Half case Left valve case 23L 23M right 23R VS7 Valve Valve Actuator valve V7 actuator V17 aC Ua Ora V27 VS8 Valve Valve V lv the actuator V8 actuator VI 8 actuator V28 VS9 Valve Valve V9 Actuator Valve Actuator VI 9 V29 Actuator VS10 Valve Valve Actuator Valve IO Actuator V20 Actuator V30 The cartridges 23L, 23M, and 23R are mounted respectively on their pumping stations PSL, PSM, PSR with their opposite sides 206 down, so that the diaphragms 212 face and couple the valve actuators. Valve actuators Vn are actuated solenoid pistons 215 (see Figure 12), which are diverted to a valve closing position. The valve actuators Vn are designed to be aligned with the valve stations of the cartridge • VSn in the manner indicated in Table 1. When a given plunger 215 is energized, the valve station of the associated cartridge is opened, allowing passage of a liquid. When the plunger 215 is not energized, the diaphragm 212 is displaced at the associated valve station, blocking the passage of liquid TL through the associated valve station. In the illustrated embodiment, as shown in Figure 12, the pumps Pl to P6 in each pumping station PSL, PSM, and PSR include the rotation of the rotors 216 of the peristaltic pump. The rotors 216 can be moved between a retracted condition (shown in Figure 13), outside the coupling with the respective tube loop, and an operation condition (shown in Figure 14), in which the rotors 216 couple the respective tube against a pump channel 218. By which the pumps Pl and P6 can be operated in three conditions: (i) in a pumping condition, during which the rotors of the pump 216 rotate and are in their operating position to attach the pump tubing to the pump channel 218 (as shown in Figure 14). The rotating pump rotors 216 therefore transport the fluid in a peristaltic manner through the pipe loop - (ii) in an open, pumping condition, during which the rotors of the pump 216 do not rotate and they are in their retracted position, so as not to couple the pump's pipe loop (as shown in Figure 13) - The open, pumping condition therefore allows the fluid to flow through the pump tube loop in absence of rotation of the pump rotor (iii) in a closed, pumping condition, during which the rotors of the pump 216 do not rotate and, the rotors of the pump are in the operating condition. pump 216 are motionless whereby they couple the loop of the pump tubing, and function as a clamp to block the flow of fluid through the pump tubing loop.Of course, equivalent combinations can be achieved under the conditions of pumping u using the rotors of the peristaltic pump that do not retract, by properly placing the clamps and the tubing paths upstream and downstream of the pump rotors. Additional structural details of the cartridges 23L, 23M, 23R, the peristaltic pumps PI to P6, and the valve actuators VI to V30 are not basic in the invention. These details are described in U.S. Patent No. 5,427,509, entitled "Peristaltic Pu b Tube Cassette with Angle Port Tube Connectors", which is incorporated herein by reference.

C. Piping for Fluid Flow The fluid circuit 200 additionally includes lengths of flexible plastic tubing, designated TI to T20 in Figure 6. Flexible tubing TI to T20 couples the cartridges 23L, 23M, and 23R to the processing vessel 14, to the external source and to the collection containers or bags, and to the blood donor / patient. The function of the fluid flow of the TI to T20 pipeline will be described later in relation to the accumulation and collection of the MNCs. The following summarizes, from a structural point of view, the union of the TI pipe to T20, as shown in Figure 6: The TI pipe extends from the donor / patient (by means of a conventional phlebotomical needle, not shown) through an external clamp C2 to channel C4 of the left cartridge 23L. The pipe T2 extends from the TI tube through an external clamp C4 to the C5 channel of the medium cartridge 23M. The pipe T3-extends from an air detection chamber DI to the channel C9 of the left cartridge 23L. "The pipe T4 extends from the drip chamber DI to the port 48 of the processing vessel 14. The pipe T5 extends from the port 50 of the processing vessel 14 to the channel C4 of the medium cartridge 23M. The pipe T6 extends from the channel C9 of the middle cartridge 23M to the connecting pipe T4 downstream of the chamber DI The pipe T7 extends from the channel 8 of the right cartridge 23R to the channel C8 of the left cartridge 23L The pipe T8 extends from the channel Cl of the 23M medium cartridge to the T7 junction pipe, T9 pipe extends from the C5 channel of the left cartridge 23L through an air detection chamber D2 and an external C3 clamp to the donor / patient (by means of a phlebotomical needle) conventional, not shown.) The UNC pipe extends from the port 46 of the processing vessel 14, through an OS optical line detector to the C4 channel of the right cartridge 23R. from channel C9 of right cartridge 23R to camera DI.

The pipe T12 extends from the channel C2 of the right cartridge 23R to a container for the purpose of receiving the platelet-poor plasma, designated co or PPP. A weight scale (not shown) detects the weight of the PPP vessel for the purpose of deriving fluid volume changes. The pipe T13 extends from the Cl channel of the right cartridge 23R to a container in order to receive the mononuclear cells, designated as MNC. The pipe T14 extends from the channel C2 of the medium cartridge 23M to a container in order to receive the red blood cells condensed from the blood, designated as PRBC. A weight balance WS detects the weight of the container of the PRBC in order to derive the volume changes of the fluid. The pipe T15 extends from a container of the anticoagulant, designated as ACD, to the channel C8 of the medium cartridge 23M. A weight scale (not shown) detects the weight of the ACD vessel for the purpose of deriving fluid volume changes. The pipe T16 and T17 extend from a container of the priming liquid, such as salt, designated as PRIME, by diverting all the cartridges 23L, 23M, and 23R, through an external clamp Cl, and intercepting, respectively, the pipe T9 (between the air detection chamber D2 and the clamp C3) and the TI pipe (upstream of the clamp C3). A weight scale (not shown) detects the weight of the PRIME vessel for the purpose of deriving fluid volume changes. The pipe T18 extends from the port 52 of the processing vessel 14 to the channel C5 of the right cartridge 23R. The pipe T19 extends from the port 54 of the processing vessel 14 to intercept the pipe T18. The pipe T20 extends from the channel C2 of the left cartridge 23L to a container in order to receive the excess waste fluid, designated as SURPLUS. A balance of weight (not shown) detects the weight of the container of the SOVEREANT for the purpose of deriving the volume changes of the fluid. The portions of the pipe are joined at the navel 30 (see Figure 1). The umbilicus 30 provides a fluid flow communication between the interior of the processing vessel 14 within the centrifugal field and other immobile components of the circuit 200 located outside the centrifugal field. Uh non-rotating holder 32 (omega zero) retains the upper portion of the navel 30 in a non-rotating position on the suspended coil and concave elements 18 and 20. A holder 34 on the rocker 22 rotates the middle portion of the navel 30 to a first (omega one) velocity on the suspended coil and concave elements 18 and 20. Another holder 36 rotates the lower end of the navel 30 at a second speed twice the speed omega one (speed omega two), in which the suspended coil and concave elements 18 and 20 also rotate. This relative rotation of the navel 30 is known to maintain unwinding, thus avoiding the need for rotary seals.

IV. Separation in the Blood Processing Chamber (Global Appreciation) Before explaining the details of the procedure by which the MNCs are collected using the container 14 and the __fluid 200 circuit, the fluid dynamics of the separation of whole blood in the compartment Processing 38 will generally be described first, with reference primarily to Figures 4 and 15 to 17. Referring first to Figure 4, anticoagulated whole blood (WB) is removed from the donor / patient and conducted in the processing compartment to through the port 48. The blood processing compartment 38 includes an inner seal 60 and 66, which forms a passage entry WB 72 which is carried in an entrance region WB 74. When WB follows a circumferential flow path in the compartment 38 on the rotary shaft 28. The side walls of the container 14 are expanded to form the profiles of the outer wall (low G) 26 of the element of b obin 18 and the inner wall (high G) 24 of the concave element 20. As shown in Figure 17, WB is separated in the centrifugal field within the blood processing compartment 38 in the condensed red blood cells of the blood (PRBC , designated by the number 96), which moves towards the high wall G 24, and platelet enriched plasma (PRP, designated by the number 98), which moves by the movement of the PRBC 96 towards the low wall G 26. An intermediate layer, called the interface (designated by the number 58), is formed between the PRBC 96 and the PRP 98. Referring again to Figure 4, the inner seal 60 also creates a collection region of PRP 76 Within the blood processing compartment 38. As further shown in Figure 17, the collection region of PRP 76 is adjacent to the entry region WB 74. The speed which the PRBC 96 establishes toward the high wall G 24 in response to centrifugal force is greater in the inlet region WB 74 than elsewhere in the blood processing compartment 38. There is also relatively more plasma volume to displace towards the lower wall G 26 in the inlet region WB 74. As a result, the speeds of the relatively larger radial plasma towards the low wall G 26 occur in the input region WB 74. These higher radial velocities towards the low wall G 26 elude the larger numbers of platelets of the PRBC 96 in the collection region by PRP closure 76. As shown in Figure 4, the inner seal 66 also forms a lower section 70 defining a collection step of the PRBC 70. An increased barrier 115 (see Figure 15) which extends into the mass of PRBC to along the high wall G 24, creating a restricted passage 114 between it and the front of the high isoradial wall G 24. The restricted passage 114 allows the PRBC 96 present along the high wall G 24 to move forward of the Pub 115 in the collection region 50 of the PRBC, to be conducted by the collection step of PRBC 78 to the port of PRBC 50. Simultaneously, the increased barrier 115 blocks the passage of the PRP 98 in front of it.

As shown in Figures 15, 16A and 16B, the high wall G 24 also projects towards the low wall G 26 to form a tapered ramp 84 in the collection region of PRP 76. The ramp 84 forms a narrow passage 90 to along the low wall G 26, along which the PRP layer 98 extends. The ramp 84 maintains the interface 58 and PRBC 96 away from the collection port of PRP 46, while allowing the PRP 98 to reach the collection port of PRP 46. In the illustrated and preferred embodiment (see Figure 16A), the ramp 84 is oriented at an angle not parallel to less than 45 ° (and preferably about 30 °) with respect to the port axis of PRP 46. The angle averages the spill over the interface and PRBC through the narrow passage 90. As shown in Figures 16A and 16B, the ramp 84 also displays the interface 26 for viewing through a side wall of the container 14 by an associated interface controller 220 (see Figure 19). The interface controller 220 controls the relative flow rates of the WB, of the PRBCs, and of PRPs through their respective ports 48, 50, and 46. In this manner, the controller "220 may maintain the interface 58 at the prescribed locations on the ramp, either near the narrow pass 90 ( as shown in Figure 16A), or spaced away from the narrow passage 90 (as shown in Figure 16B) By controlling the position of the interface 58 on the ramp 84 relative to the narrow passage 90, the controller 220 can also control the platelet content of the plasma collected through port 46. The concentration of platelets in the plasma increases with proximity to interface 58. Maintaining interface 58 at a relatively low position on ramp 84 (as shown in FIG. Figure 16B), the platelet enriched region is kept away from port 46, and the plasma conducted by port 46 has a relatively low platelet content, keeping interface 58 in a high position on ramp 84 (as shown in FIG. shows in Figure 1 6A), closer to port 46, the plasma conducted by port 46 is enriched with platelets. Alternatively, or in combination, the controller could control the location of interface 58 by varying the speed at which WB was introduced into the blood processing compartment 38, the speed at which PRBC was transported from the blood compartment. blood processing 134, or both.

Additional details of the preferred mode for the interface controller are. describe in US Patent 5,316,667, which was incorporated herein by reference. As shown in Figure 15, the radially opposed surfaces 88 and 104 form a flow restriction region 108 along the high wall G 24 of the entrance region of the WB 74. As also shown in Figure 17 , the region 108 restricts the flow of the WB in the inlet region of the WB 74 at a reduced rate, whereby uniform perfusion of the WB is caused in the blood processing compartment 38 along the wall low G 26. This uniform perfusion of the WB occurs adjacent to the collection region of PRP 76 and in a plane that is approximately equal to the plane at which the preferred and controlled position of the interface 58 remains. Once in the region narrow 108 of dam 104, the PRBC 96 rapidly moves toward the high wall G 24 in response to the centrifugal force. The narrow region 108 leads WB approximately to the preferred and controlled height of the interface 58 in the input region 74. The WB carried down or above the controlled height of the interface 58 58 in the input region 74 will immediately seek the height of the interface and, thus, oscillate on it, causing unwanted secondary flows and disturbances along the interface 58. By bringing the WB approximately to the level of the interface in the input region 74, the region 108 reduces the incidence of secondary flows and disturbances along the interface 58. As shown in Figure 15, the low wall G 26 tapered out away from the axis of rotation 28 towards the high wall G 24 in the flow direction of the WB , while the front of the high wall G 24 retains a constant radius. The taper can be continuous (as shown in Figure 15) or may occur in the form of a step. These contours along the high G and low G 24 and 26 walls produce a dynamic condition of the circumferential plasma flow generally transverse to the "field of centrifugal force in the direction of the collection region of PRP 76. As schematically detailed in FIG. Figure 18, the condition of the circumferential plasma flow in this direction (arrow 214) continually pulls the interface 58 back into the collection region of PRP 76, where the highly radial plasma flow conditions already described, exist to collect even more The platelets of the interface 58. Simultaneously, the counterflow models serve to circulate again the other heavier components of the interface 58 (the lymphocytes, monocytes, and granulocytes) in the mass of the PRBC, away from the PRP flow. Within this dynamic condition of the circumferential plasma flow, MNCs (designated as in Figure 18) are initially established along e the high wall G 24, however eventually floats to the surface of the interface 58 near the high hematocrite PRBC collection region 50. The taper of the low wall G creates the backflow patterns of the plasma, shown by the arrows 214 in Figure 18. These counterflow models 214 extract the MNC back to the collection region of low hematocrit PRP 76. The MNCs regroup again near the collection region of low hematocrit PRP 76 towards the high wall G 24. MNC circulate in this path, designated 216 in Figure 18, while the WB is separated in the PRBC and PRP. The MNCs are thus collected and "stationed" on this path 216 confined within compartment 38 away from the collection region of the PRBC 50 and the collection region of PRP 76.

Additional details of the separation dynamics in the processing compartment 38 are found in U.S. Patent 5,573,678, which is incorporated herein by reference.

V. Mononuclear Cell Process Procedure The centrifuge 10 includes a process controller 222 (see Figure 19) which commands the operation of the fluid circuit 200 to carry out the collection of the prescribed MNCs and the collection procedure 224 using the container 14. As shown in Figure 20, the method 224 comprises a preprocessing priming cycle 226 which primes the fluid circuit 200. The following procedure 224 includes a preliminary processing cycle 228, which processes the PPP of the whole blood obtained from the donor / patient for later use in procedure 224 as a means of suspension for the collected MNCs. The following procedure 224 includes at least one cycle of the main processing 230. The main processing cycle 230 comprises a collection stage 232, followed by a collection stage 234.

Collection step 232 includes a series of collection steps 236 and 238, during which whole blood is processed to accumulate the mononuclear cells in the first compartment 38, in the manner previously described. The collection stage as well includes a series of collection phases 240, 242, 244, and 246, during which the accumulation of mononuclear cells is transferred from the first compartment 38 into a collection container of the MNCs coupled to the 200 circuit. it adds to the MNC, the suspension medium collected during the preliminary processing cycle 228. Normally, the main processing cycle 230 will be carried out more than once during a given procedure 224. The number of processing cycles 230 conducted in a given procedure 224 will depend on the total volume of the MNC sought to be collected. For example, in a representative procedure 224, five cycles of the main processing 230 are repeated, one after the other. It can be processed during each cycle of the main processing 230, from about 1500 to about 3000 ml of whole blood, to obtain a volume of the MNCs per cycle of about 3 ml. At the end of the five processing cycles 230, an MNC volume of approximately 15 ml can be collected, which is suspended in a final dilution of the PPP of approximately 200 ml.

A. Preprocessing Priming / Ballast Sequence Before a donor / patient is coupled to the fluid circuit 200 (via line TI and T9), the controller 222 conducts a priming cycle 228. During the primer cycle 228, the controller 222 commands the centrifuge 10 to rotate the coil and concave elements 18 and 20 on the axis 28, while arranging the pumps Pl to P6 to transport a sterile priming liquid, such as saline, from the PRIME container and the anticoagulant from the ACD container to The length of the complete fluid circuit 15 and of the container 14. The priming liquid displaces air from the circuit 15 and from the container 14. The second compartment 40 is aided by a single pipe T18 and therefore has, in effect, a single port. of access. To achieve the priming, the compartment 40 is isolated from the flow communication with the priming liquid, while the pump PS is operated to extract air from the compartment 40, whereby a negative pressure (vacuum) condition is created in the compartment. 40. On the air deposition of the compartment 40, the communication is then opened to the flow of the priming liquid, which is removed from the compartment 40 by vacuum. The PS pump is also operated to assist in the conduction of the liquid in the compartment 40 and create a positive pressure condition in the compartment 40. The controller 222 retains the priming liquid in the second compartment 40, to counterbalance the first compartment 38 during the processing of the blood. It must, of course, be appreciated that this vacuum priming procedure is applicable to the priming of any container virtually assisted by a single access port or its equivalent.

B. Preliminary Processing Cycle The MNC collected in the MNC container is preferably suspended - in a low platelet plasma (PPP) obtained through the donor / patient's MNC. During the preliminary processing cycle 228, the controller 222 configures the fluid circuit 222 to collect a volume of the PPP preset from the donor / patient for retention in the PPP container. This volume is then used as a suspension medium for the MNCs during processing, as well as being added to the MNCs after processing to achieve the desired final dilution volume. Once the donor / patient has phlebotomized, the controller 222 configures the pump stations PSL, PSM, and PSR to initiate the preliminary processing cycle 228. As previously described, during this cycle 228, whole blood is centrifugally separated in compartment 38 in condensed red blood cells (PRBC) and platelet-enriched plasma (PRP). PRBCs are returned to the donor / patient, while mononuclear cells accumulate in compartment 38. When MNCs accumulate in compartment 38, a portion of the separated plasma component is removed and collected to be used as a suspension medium. the MNC. During this cycle 228, the controller 222 maintains the interface 58 at a relatively low position on the ramp 84 (as detailed in Figure 16B). As a result, the plasma that is transported from the compartment 38 and stored in the PPP container is relatively poor in platelets, and thus can be characterized as PPP. The remainder of the PPP conducted from compartment 38 is returned to the donor / patient during cycle 228. The configuration of fluid circuit 200 during the preliminary processing cycle 22B is shown in Figure 21, and is further summarized in Table 2.

Table 2: Preliminary Processing Cycle Where: • indicates an occluded pipe or closed condition. or indicates a non-occluded pipe or open condition. "indicates a pumping condition, during which the pump rotors rotate and couple the pump tubing to conduct the fluid in a peristaltic manner; or indicate an open, pumping condition, during which the pump rotors they do not rotate and do not couple to the pump's pipe loop, and therefore allow the fluid to flow through the pump's pipe loop .. I * • indicates a closed, pumping condition during which the pump's rotors do not They rotate, and mate with the pump's pipe loop, and therefore do not allow the fluid to flow through the pump's pipe loop.During the preliminary cycle 2287 the pump P2 extracts the "whole blood (WB) from the donor / patient through the TI tubing in the left cartridge 23L, in the T3 tubing, through the DI chamber, and "in the blood processing compartment 38 through the tubing T4. ACD anticoagulant through the T1 pipe 5, in the medium cartridge 23M and in the T2 pipe, to mix with the whole blood. Anticoagulated whole blood is conducted in compartment 38 through port 48. Whole blood is separated into PRP, PRBC, and the interface (including MNCs), as previously described.

The port 50 conducts the PRBCs 96 of the blood processing compartment 38, through the pipe T5 in the medium cartridge 23M. The PRBCs enter the T7 pipe through the T8 pipe, to return the donor / patient through the left cartridge 23L and the T9 pipe. Port 46 conducts the PPP from the blood processing compartment 38. The PPP continues in the UNC pipe in the right cartridge 23 R. The pump P5 transports a portion of the PPP in the T7 line to return with the PRBCs to the donor. Patient-The interface controller 220 indicates the pump flow rate P5 to maintain the interface in a low position on the ramp 84 (as shown in Figure 16B), whereby the concentration of conducted platelets of the patient is minimized. compartment 38 during this cycle Pump P6 conducts a portion of the PPP through pipe T1.2 in the PPP container, until the prescribed volume is collected for the suspension of the MNCs and the final dilution. V0Lsus.

C. Main Processing Cycle 1. Mononuclear Cell Collection Stage (MNC) (i) MNC Accumulation Phase The control 222 is now activated for the collection stage of the MNC 232 of the main processing cycle 230. First , the controller 222 configures the fluid circuit 200 for the accumulation phase of the MNC 236. For the phase 236, the controller 222 changes the configuration of the pumping station PSR to stop the collection of the PPP. controller of the interface 220 to maintain a flow rate of the pump P5 to maintain the interface at a "higher location on the ramp 84 (as shown in Figure 16A), whereby separation of the PRP is enabled. . Due to the changed combination, the pump P6 also recirculates a portion of the PRP to the blood processing chamber 38 to improve the efficiency of the platelet separation, as will be described later in greater detail. The configuration for the accumulation phase of the MNC 236 of the collection stage of the MNC 232 is shown in Figure 22 and is further summarized in Table 3. Table 3: Mononuclear Cell Collection Condition (Accumulation Phase) of the MNC) Where: • indicates an occluded pipe or. closed condition. or indicates a non-occluded pipe or open condition. ^ indicates a pumping condition, during which the rotors of the pump rotate and couple the pump tubing to conduct the fluid in a peristaltic manner.

It indicates an open, pumping condition, during which the pump's rotors do not rotate and do not couple to the pump's pipe loop, and therefore allow the fluid to flow through the pump's pipe loop. 1 ^ indicates a closed, pumping condition during which the rotors of the pump do not rotate, and are coupled with the pipe loop "of" the pump, and therefore do not allow the fluid to flow through the loop of the pump. pump tubing. 1. Efficiencies that Promote High Separation of Platelets by Recirculating PRB Normally, platelets are not collected during an MNC procedure. Instead, it is desired that they return to the donor / patient. A high MPV main platelet volume (expressed in femtoliters, fl, or cubic microns) is desired for the separated platelets, when an efficient high platelet separation is denoted. MPV can be measured from a PRP sample by conventional techniques. Major platelets (ie, greater than about 20 fentoliters) are more likely to be caught at the 58 interface and not between the PRP for return to the donor / patient. This will produce a reduced population of larger platelets in the PRP, and therefore a lower MPV, to return to the donor / patient. The establishment of radial conditions of plasma flow sufficient to elevate the major platelets of the interface 58, as previously described is highly dependent on the input hematocrit Hi of the WB entering the blood processing compartment 38. For this reason, the pump 6 recirculates a portion of the PRP flowing in the UNC pipe again into the inlet port of the WB 48. The recirculation of the PRP flows through the right cartridge 23R in the Til pipe, which joins the pipe T4 coupled to the inlet port 48. The recirculation of the mixture of PRP with WB entering the blood processing compartment 38, whereby the input hematocrit Hi is low. The controller indicates the "recirculation flow rate" of the PRP Qrecirc of pump P6 to achieve a desired input hematocrit Hi. In a preferred application, H is not greater than about 40%, and, preferably, about 32%, which will achieve a high MPV. The input hematocrit Hi can be measured conventionally by an on-line detector in line T4 (not shown). The input hematocrit Hx can be determined empirically based on the direction of the flow conditions, as described in copending US Patent Application Serial No. 08 / 471,883, which is incorporated therein by reference. 2. Promotion of the High Concentration of the MNC and Purity by Recirculation of the PRBC Co or was schematically described in the Figure 18, the plasma counterflow (arrows 214) in compartment 38 dredges interface 58 back to the collection region of PRP 76, where improvement of radial plasma flow conditions collect platelets outside interface 58 to return them to the donor / patient. In the flow counter models 214, other heavier components of the interface 58 also circulate, such as lymphocytes, monocytes, and granulocytes, again for circulation in the PRBC mass. Meanwhile, due to the relatively high hematocrit in the collection region of the PRBC 80, the MNC float near the 80 region to the surface of the interface 58. There, the MNCs are extracted by the counterflow of the plasma 214 to the region of collection of PRP of low hematocrit 76. Due to the low hematocrit in this region 76, the MNCs regroup again towards the high wall G 24. Arrow 216 in Figure 18 shows the desired circulating flow of the MNC when it accumulates in the compartment 38. It is important to maintain a desired output HBC PRBC hematocrit in the collection region of PRBC 50. If the output hematocrit H0 of the PRBC falls below a low threshold value provided (eg, at about 60% low), "most MNCs will not" circulate as a cell mass, as shown by arrow 216 in Figure 18. Exposed to a low H0, all or some of the MNCs will fall to float to interface 58. Instead, the MNCs will remain banded along the high wall G and will be carried out of compartment 38 with the PRBCs. An insufficient result provides the MNCs. On the other hand, if H0 exceeds a given high threshold value (for example, approximately 85%), the larger numbers of the heavier granulocytes will float at interface 58. As a result, less "granulocytes will be taken away from the 58 interface to return the PRBCs to the donor / patient, whereas more granulocytes will occupy the 58 interface and contaminate the MNCs.

For this reason, during the collection stage of MNC 232, process controller 222 commands pump P4 to recirculate one. portion of the PRBCs that flow into the T5 pipe again at the inlet port of the WB 48. As shown in Figures 21 and 22, the recirculation of the PRBCs flows through the medium cartridge 23M into the T6 pipe, which is attached to the tubing T4 coupled to the inlet port 48. The recirculation of the mixture of the PRBCs with the WB enters the blood processing compartment 38. Generally speaking, the magnitude of the output hematocrit H0 varies reciprocally as a function of the recirculation flow rate of the PRBC Qr, which is governed by the pump P4 '(PRBC) and the pump P2 (WB). By providing a flow rate for the WB set by the pump P2, the hematocrit H0 de-output can be increased by lowering Qr, and, reciprocally, the output HQ hematocrit can be decreased by raising Qr. The exact relationship between Qr and Hc takes into account the centrifugal acceleration of the fluid in the compartment 38 (governed by the magnitude of the centrifugal forces in the compartment 38), the compartment area 38, as well as the velocity of the inlet flow of the compartment. Whole blood (Qb) in the compartment 38 (governed by the pump P2) and the output flow rate of PRP (Qp) from the compartment 38 (governed by the interface control pump P5). There are several ways to express this relationship and therefore quantify Qr based on a desired H, D. In the illustrated embodiment, the controller 222 periodically tests Qb / QP, and Qr- In addition the active centrifugal force factors are taken into account in the compartment 38, the controller derives a new recirculation pumping speed of the PRBC Qr (NEW) for pump P4, based on an indication of H0, as follows: (i) start at a sample time n = 0 (ii) Calculate current Qr as follows: _r- [«P - < > "• _ _ -I - ui: *?:! where: H0 is the indicated output hematocrit value, expressed as a decimal (for example, 0.75 for 75%). a is the acceleration of the fluid, governed by the centrifugal forces, calculated as follows: 7S? a = where O is the speed of rotation of compartment 38, expressed in radiants per second. r is the radius of gyration. g is the unit of gravity, equal to 981 cm / sec2. ~ A is the compartment area 38. k is the constant hematocrit and m is a constant separation realization, which is derived based on the empirical data and / or the theoretical model. In the preferred embodiment, the following theoretical model is used: ß ß H¿ H0 (1 - H0) * + 1 = a A CR where CH = 1.08 Sr and where: ß is a sensitive term of shear stress as defined: ß + y where: < based on the empirical data, b = 6.0 s ~ n and n = 0.75, and the shear velocity is defined as: t = du / dy where (u) is the flow velocity e (y) is a spatial dimension. and where: Sr is the sedimentation factor _ of the empirically derived red blood cells, which, in the empirical data, can be placed at 95 x 10 ~ 9s.This model is based on the * Equation (19) of Brown, " The Physics of Continuous Flow Centrifugal Cell Separation ", Artificial Organs; 13 (l): 4-20, Raven Press, Ltd., New York (1989) (The" Brown Article "), which is incorporated herein by reference The plot of the model appears in Figure 9 of Brown's article.The previous model is linearized using a simple linear regression over a range of practical operation, assumption of the process conditions of the blood.Algebraic substitutions are made based on the expressions following: where: Qo is the flow velocity of the PRBCs through the outlet pipe T5, which "can be expressed as: This linearization provides a simplified curve in which the value of (m) constitutes the slope and the value d.e (k) constitutes the intersection y. In the simplified curve, the slope () is expressed as follows: ß m = 338. 3 ( where: ß / Sr can be based on empirical data, expressed as a constant value of 1.57 / μs. Therefore, in the simplified curve, m has a value of 531.13. A range of values for m between approximately 500 and approximately 600 is generally believed to be applicable to the "centrifugal, the continuous flow of the whole blood separation procedures." For the simplified curve, the value of the intersection y for (k) equals 0.9489. A range of values for k between about 0.85 and about 1.0 is believed to be generally applicable to centrifugal whole-blood separation procedures. (iii) Average calculation Qr Qr is measured in the intervals selected, and these instantaneous measurements are averaged over the process period, as follows: Qr (PROMED I 0; = [0 .95 (Qr (PROMED I OULTIMC J + [0. 05 * Qr] iv) New calculation Qr, as follows QS (NEW) = Qr (AVERAGE) * F where: F is an optional CONTROL factor, which 'enables Qr control (when F = 1), or disables Qr control (when F = 0), or enables a Qr scale based on system variations (when F is expressed as a fraction between 0 and 1). F may comprise a constant or, alternatively, may vary as a function of the processing time, for example, by starting at a first value at the output of a given procedure and changing to a second or more values as the progress of the procedure. (v) Keep Qr within the prescribed limits (for example, between 0 ml / min and 20 ml / min) SI Qr (NEW) > 20 ml / min THEN Qr (NEW) = 20 ml / min ENDIF IF Qr (NEW) < 0 ml / min THEN Qr (NEW) = 0 ml / min ENDI F n = n + 1 During the collection stage of the MNC 232 (Figure 22), the controller 222 simultaneously maintains and indicates the multiple flow velocities of the pump to achieve optimum processing conditions in compartment 38 for the accumulation of a high yield of high purity MNCs. The controller indicates and maintains the input flow velocity Qb of the WB (by means of the pump P2), the output flow rate QP of PRP (by means of the pump P5), the velocity Qp.ecirc of flow of recirculation of PRP (by means of pump P6), and the speed Qr_of recirculation flow of PRBC (by means of pump P4). By providing a flow rate Qb of the WB input, which is normally indicated for the comfort of the donor / patient and the achievement of an acceptable process time, the controller 222: (i) commands the P5 pump to maintain a Qp indicated for retaining a position of the desired interface on the ramp 84, and in such a way that it achieves the platelet concentrations desired in the plasma (PPP or PRP); (ii) commands the P6 pump to maintain a QRecirc indicated to retain the desired Hi-input hematocrit (e.g., between about 32% and 34%), and thereby achieve the high platelet separation efficiencies; and (iii) command pump P4 to maintain a Qr indicated to hold a. desired H0 output hematocrit (eg, between about 75% to 85%), and in such a way as to prevent granulocyte contamination and maximize MNC yields. (ii). Second Phase (Collection of the PRBC) The .controller 222 ends the accumulation phase of the MNC 236 when a pre-set volume of whole blood is processed (for example, 1500 ml to 3000 ml). Alternatively, the accumulation phase "" "of the MNC may end when a designated volume of the MNC is collected.The controller 222" then enters the collection phase of the PRBC 238 of the collection stage of the MNC 232. In in this phase 238, the configuration of the PSM pump station is altered to stop the return of the PRBCs to the donor / patient (closing V14), stopping the recirculation of the PRBCs (closing the valve V18 and placing the pump P4 in a condition of pumping, closed, and in turn driving the PRBCs to the PRBC container (opening V15) This new configuration is shown in Figure 23, and is further summarized in Table 4.

Table 4: Collection stage of the mononuclear cell (Collection phase of the PRBC) Table: Mononuclear Cell Collection Stage (Collection Phase of the PRBC) (Continued) where: • indicates an occluded pipe or closed condition. indicates a pipeline not occluded open condition. ™ indicates a pumping condition, during which the rotors of the pump rotate and couple the pump tubing to conduct the fluid in a peristaltic manner. If or indicates a pumping condition, open, during which the pump rotors do not rotate and do not couple to the pump's pipe loop, and therefore allow the fluid to flow - through the pump's pipe loop. YES • indicates a closed, pumping condition, during which the pump's rotors do not rotate, and are coupled with the pump's pipe loop, and therefore does not allow the fluid to flow through the pipe loop of the pump. bomb. In this phase 238, the PRBCs on the line T5 are conducted through the medium cartridge 23M, in the line operated in this phase 238 to a desired volume of the PRBCs (eg, 35 ml to 50 ml) collected in the container the PRBC. This volume of the PRBCs is then used in the withdrawal phase of the MNCs 240 from the collection stage of the MNCs 234, as will be described later in greater detail. The controller 222 ends the collection phase of the PRBC 238 which detects (gravimetrically, using a WS weight balance) the container of the PRBCs that retains the desired volume of the PRBCs. The completion of the collection stage of the MNC 232 of the main processing cycle 230. 2. Mononuclear Cell Collection Stage (i) First Phase (MNC Removal) The controller 222 enters the collection stage of the MNC 234 of the main processing cycle 230. In the first phase 240 of this stage 234, the blood The whole sample is withdrawn and recirculated back to the donor / patient without passing through the blood processing compartment 38. The PRBCs collected in the container of the PRBCs in the collection phase of the PRBC preceded 238 are returned to the processing compartment 38 to through the inlet pipe of the WB T4, while turning or the compartment 38 continues. The MNCs accumulated in the compartment 38 during the collection stage of the MNC 232 are conducted with the PRP through the pipe UNC outside the compartment 38. The configuration of the fluid circuit 15 during the phase of removal of the MNCs 240 of the stage of collection of MNCs 234 is shown in Figure 24A, and is further summarized in Table 5: Table 5: Mononuclear Cell Collection Stage (MNC Withdrawal Phase) where: • indicates an occluded or closed condition pipe. or indicates an unoccluded or open condition pipe. ^ indicates a pumping condition, during which the rotors of the pump rotate and couple the pump tubing to conduct the fluid in a peristaltic manner. YES o indicates a pumping condition, open, during which the pump rotors do not rotate and do not couple to the pump pipe loop, and therefore allow the fluid to flow through the pump pipe loop. YES • indicates a closed, pumping condition, during which the pump's rotors do not rotate and mate with the pump's pipe loop, and therefore do not allow the fluid to flow through the pump's pipe loop. As shown in Figure 24A, the controller 222 closes the outlet pipe of the PRBC T5 while the PRBCs are driven by the pump P4 from the PRBC container through the pipes T14 and T6 in the pipe T4, for the introduction into the compartment 38 through the input port of the WB 48. The controller 222 starts a cycle against time in TCYCIHICIO. In PRBC flow from the PRBC container through the entry port of WB 48 the hematocrit increases in the collection region of PRP 76. In response, the concentrated region of the MNCs accumulated in compartment 38 (as shown in Figure 18), floats to the surface of interface 58. The volume of the PRBCs shifts the PRP through the PRP 46 exit port. interface 58, and with this, the concentrated region of the MNCs (designated MNC Region in Figure 24A) are also moved out of compartment 38 through the PRP 46 exit port. The MNC Region moves as length of the PRP UNC pipe to the optical detector OS. As shown in Figure 28, within the UNC pipeline, a PRP region 112 precedes the Concentrated Region of the MNCs. The PRP in this region 112 is conducted in the PPP vessel through the right cartridge 23R and the pipe T12 '(as shown in Figure 24A). A PRBC region 114 also follows the concentrated Region of the MNC within the TÍO pipeline. A first transition region 116 exists between the PRP region 112 and the concentrated Region of the MNCs. The first transition region 116 consists of a stable decreasing concentration of platelets (shown by a box model in Figure 28) and a stable increasing number of the MNCs (shown by a texturized model in Figure 28). A second transition region 118 exists between the Concentrated Region of the MNCs and the PRBC region 114. The second transition region 118 consists of a stable decreasing concentration of the MNCs (shown by the texturized model in Figure 28) and a stable increasing number of the PRBCs (shown by a wave model in Figure 28). Seen by the optical detector OS, the regions 112 and 116 precede the Region of the MNC and the regions 118 and 114 that carry the Region of the MNC present in optical transition densities in which the Region of the MNC can be discerned. The OS optical detector detects changes in. the optical density in the liquid conducted by the UNC pipe between the PRP output port 46 and the right cartridge 23R. As shown in Figure 28, the optical density will change from a low value, which_ indicates a highly transmissible light (i.e. ', in the PRP 112 region), to a high value, which indicates a highly absorbent light (i.e. , in the region of the PRBC 114), when the MNC Region to pass the OS optical detector.

In the embodiment illustrated and shown in Figure 28, the optical detector OS is a conventional hemoglobin detector, used, for example, in the Autopheresis-C blood processing device sold by Fenwal Division of Baxter Healthcare Corporation. The detector OS comprises a red light that emits diode 102, which emits light through the UNC pipe. Of course, other wavelengths could be used, such as green or infrared. The OS detector also includes a PIN 106 detector on the opposite side of the UNCLE pipe. The controller 222 includes a processing element 100, which analyzes the voltage signals received from the emitter 102 and the detector 106 to calculate the optical transmission of the line in the UNC pipe which is called OPTTRANS. Various algorithms can be used by the processing element 100 to calculate OPTTRANS. For example, OPTRANS can match the output of the diode detector 106 when the red light emitted by the diode 102 is on and the liquid flows through the UNCLE (RED) pipe. The optical background "noise" can be filtered from RED to obtain OPTTRANS, as follows: COR (SPILL RED) OPTTRANS = CORRREF where COR (SPILL RED) is calculated as follows COR (SPILL RED) = RED-REDBKGRD where: RED is the output of the diode detector 106 when the diode 102 emits red light is on and the liquid flows through the pipe UNCLE; and where CORREF is calculated as follows: CORREF = REF-REFBKGRD where: REF is the output of diode 102 that emits red light when the diode is on; and REFBKGRD is the output of the diode 102 that emits light r "ja 1 when the diode is off.The processing element 100 normalizes the OS detector to the optical density of the PRP of the donor / patient, obtaining data from the OS detector during the MNC collection preceded 232, when the PRP of the donor / patient is conducted through the UNC pipe.This data establishes a linear base optical transmission value for the pipeline and the donor / patient PRP (OPTTRANSBASE) • For example , OPTTRANSBASE can be measured at a selected time during collection step 232, eg, equidistant through step 232, using any filtering or unfiltered detection scheme, as described above.Alternatively, a set of transmission values optics are calculated during the collection stage of the MNC 232 using any filtered or unfiltered detection scheme.The group of values are averaged over the complete collection stage to derive OPTTRANSBASE. The processing element 100 continues during the phase of withdrawal of the subsequent MNC 240 to detect one or more optical transmission values for the UNC pipe and the liquid flowing therein.

(OPTTRANSRECOLECTION) during the phase of removing the MNCs, 240. OPTTRANSRECOLECTION may comprise a single reading detected at a selected time of the withdrawal phase of MNCs 240 (eg, equidistant through phase 240), or may comprise an average of multiple readings taken during the phase of MNC removal 240 The processing element 100 derives a normalized value of DENSITY by setting OPTTRANSBASE to 0.0, setting the optical saturation value to 1.0, and setting the value of OPTTRANSRECO ECTION "proportionally in the normalized 0.0 to 1.0 value range." As shown in the Figure 28, the processing element 100 retains two predetermined threshold values THRESH (l) and THRESH (2). The value of THRESH (l) "corresponds to a nominal value- selected by the DENSITY (for example, 0.45 on a standardized scale from 0.0 to 1.0), which has been empirically determined to occur when the concentration of the MNCs in the first transition region 116 meets a purpose of the preselected process.The value "of THRESH (2) corresponds to another nominal value selected by ___ DENSITY (for example, 0.85 on a normalized scale from 0.0 to 1.0), which has been empirically determined to occur when the concentration of the PRBCs in the second transition region 118 exceeds the purpose of the preselected process. The liquid volume of the UNC pipe between the optical detector OS and the valve station V24 in the right cartridge 23R constitutes a known value, which is input to the controller 222 as a first volume equivalent to VOLOFF.I. calculates a first value of the time control based on VOLO Fu and the pumping speed of the pump P4 (QP), as follows: VOL, OF? - (I) Ti mpoa - x 60 ¿B In the illustrated and preferred embodiment, the operator can specify and deliver to the controller 222 a second equivalent volume VOL0FF_U '.. which represents a nominal additional volume (shown in Figure 28) to increase the total collected volume of the MNC VOLMNC. L amount of V0LOFF.2. takes into account the system and variants of the processing, as well as the variants between the donors / patients in the purity of the MNCs. The controller 222 calculates a second value of the time control Time2 based on V0L0FF.2. and the speed of pump P4 (Qp4), as follows: VOLOFF Í (ZZZ :) Time2 x 60 As the pump operator P4 conducts the PRBCs through the input port WB 48, the interface 58 and the MNC Region advance through the PRP UNC pipe to the optical detector OS. The PRP that precedes the MNC Region advances beyond the OD optical detector, through the T12 pipe, and into the PPP vessel. When the MNC Region reaches the OS optical detector, the OS detector will detect the DENSITY THRESH (l). In this event, controller 222 initiates a first timer TCi. When the optical detector OS detects the DENSITY = THRESH (2) the controller 222 initiates a second timer TC_. The detected MNC volume can be derived based on the interval between TCX and TC2 for a given QP4. As time progresses, the controller 222 compares the magnitudes of TCi for the first time control Ti, as well as compares TC2 to the second time control T2. When TC = Ti, the leading edge of the indicated Region of the MNC has arrived at the valve station V24, as shown in Figure 248. The controller 222 controls the V24 station to open it, and controls the valve station V25 for close. The controller 222 marks this event in the cycle against time as TCYCS ITCH. The indicated MNC Region is conducted on the T13 pipe that directs the MNC container. When TC2 = T ^, the second volume equal in VOLOFFÍ) has also been conducted in pipe T13, as shown in Figure 24C. The total volume of the MNC selected for collection (VOLMNC) during the given cycle is therefore present in the T13 pipe. When TC =?, Controller 222 controls pump P4 to stop it. In addition, the advance of VOLMNC in the pipe T13 consequently ceases. The controller 222 derives the volume of PRP that was conducted in the PPP vessel during the withdrawal phase of the preceded MNCs. This volume of PRP is derived (which is "designated VOLPRP), as follows: TCYC SWITCH - TCYCr VOLpRp = In a preferred embodiment, controller 222 ends the MNC removal phase, independent of TCi and TC2 when pump P4 conducts plus a specified fluid volume of PRBCs after TCYCINITION (eg, more than 60 ml). This time-out circumstance occurs, for example, if the optical detector OS does not detect THRESH (l). In this time-out volumetric circumstance, VOLPRP = 60 - VOLOFF.U ^ Alternatively, or in combination with a volumetric timeout, the controller 222 can complete the MNC removal phase independent of TCi and TC2 when the weight balance WS for the PRBC container, it detects a weight less than a prescribed value (for example, less than 4 grams, or the equivalent weight of a fluid volume less than 4 ml). (ii) Second Phase (PRP flow) Once the MNC Region is positioned as shown in. Figure 24C, the controller • .222 enters the PRP flow phase 242 of the collection stage of the MNC 234. During this phase 242, the controller 222 configures the circuit 200 to move the VOLPRP out of the PPP container and the pipe T12 and in the blood processing compartment 38. The configuration of the fluid circuit 200 during the flow phase of PRP 242 is shown in Figure 25, and is summarized further in Table 6.

Table 6: Mononuclear Cell Collection Stage Table 6: Mononuclear Cell Collection Stage (PRP Flow Phase) (Continued) Where: • indicates an occluded or closed condition pipe, or indicates a non-occluded or open-condition pipe. • ^ indicates a pumping condition, during which the rotors of the pump rotate and couple the pump tubing to conduct the fluid in a peristaltic manner. YES o indicates a pumping condition, open, during which the pump's rotors do not rotate and do not couple to the pump's pipe loop, and therefore allow the fluid to flow through the pump's pipe loop.

YES • indicates a pumping condition, closed, during which the pump's rotors do not rotate and in which they are coupled with the pump's pipe loop, and therefore do not allow the fluid to flow through the pipe loop. the bomb. During the PRP flow phase 242, the controller 222 configures the pumping stations PSL, PSM, and PSR to stop the recirculation of whole blood, and, while continuing to turn the compartment 38, to pump _ -_VOLPRP to the processing compartment 38 through the Til pipe. The VOLPRP is driven by the pump P6 through the pipe T12 in the right cartridge 23R, and from there to the pipe Til, to enter the processing compartment 38 through the pipe T4 and the port 48. The PRBCs are they lead from the processing compartment 38 through the port 50 and the pipe T5 in the "middle cartridge 23M, and from there in the pipes T8 and T7 in the left cartridge 23L.The PRBCs are conducted in the pipe T9 to return it to the donor / patient No other fluid is transported in the fluid circuit 15 during this phase 242. The return of V0LPRP restores the volume of liquid in the PPP to VOLsus container as it was collected during the preliminary process cycle 228 previously described. The return of VOLPcP also preserves a low platelet population in the VOLSus in the sealed PPP container for the suspension of the MNCs.The return of VOLPRP also conducts the MNC residues in the first tract region. position 116 before TCi = Ti (and therefore not part of the VOLMNC). returning to the processing compartment 38 for further collection in a subsequent main process cycle 230. (iii) Third Phase (MNC Suspension) With the return of the _V0LPRP to the compartment 38, the controller 222 enters the phase of suspension of the -MNC 244 of the collection stage of the MNC 234.

During this phase 244, a portion of the VOLSUs in the PPP container is driven with the VOLMNC in the MNC container. The configuration of the fluid circuit 200 during the suspension phase of the MNCs 244 is shown in Figure 26, and is further summarized in Table 7.

Table 7: Collection stage of the mononuclear cell Where: • indicates an occluded or closed condition pipe. or indicates an unoccluded or open condition pipe. ™ indicates a pumping condition, during which the rotors of the pump rotate and couple the pump tubing to conduct the fluid in a peristaltic manner.

[LS-? I or indicates a pumping condition, open, during which the pump rotors do not rotate and do not couple to the pump pipe loop, and therefore allow the fluid to flow through the pipe loop of the pump. bomb. Ll • indicates a closed, pumping condition, during which the pump's rotors do not rotate and in which they are coupled with the pump's pipe loop, and therefore do not allow the fluid to flow through the pipe loop. the bomb. In the phase of suspension of the MNC 244, the controller closes the C3 to stop the return of the PRBC to the donor / patient. A predetermined aliquot of VOLSus (eg 5 ml to 10 ml) is conducted by pump P6 through line T12 into right cartridge 23R and then into line T13. As shown in Figure 26, the VOLsus aliquot further advances the VOLMNC through the T13 pipe in the MNC vessel. (iii) Fourth Phase (Cleaning) At this time, the controller 222 enters the cleaning phase, final 246 of the collection stage of the MNC 234. During this phase 246, the controller 222 returns the PRBC residing in the pipe UNCLE to the processing compartment 38. The configuration of the fluid circuit 200 during the cleaning phase 246 is shown in Figure 27, and is further summarized in Table 7.

Table 7: Mononuclear Cell Collection Stage (Cleaning phase) Where: • indicates an occluded or closed condition pipe. indicates an unoccluded or open condition pipe. "indicates a pumping condition, during which the pump rotors rotate and couple the pump tubing to drive the fluid in a peristaltic manner, SI or indicates an open, pumping condition, during which the pump rotors they do not rotate and do not couple to the pump's pipe loop, and therefore allow the fluid to flow through the pump's pipe loop .. I • indicates a closed, pump condition during which the pump's rotors do not rotate and in which they are coupled with the pump pipe loop, and therefore do not allow the fluid to flow through the pipe loop of the pump, the cleaning phase 246 returns any residue of the MNC present in the second region. of transition 118 (see Figure 28) then TC2 = T2 (and therefore not part of the VOLSEN) / returning to the processing compartment 38 for the additional collection of a subsequent processing cycle.In the cleaning phase 246, the Ontroller _ 222 closes all valve stations in the left and middle cartridges 23L and 23M and configures the right pump station PSR for the PRBCs circulated from the UNC pipe back into the processing compartment 38 through the Til and T4 pipes . During this period, no component has been extracted from or returned to the donor / patient. At the end of the cleaning phase 246, the controller 222 begins a new cycle of the main processing 230. The controller 222 repeats a series of cycles of the main processing 230 to the desired volume of the MNCs indicated for the full procedure to be reached. At the end of the last cycle of the main processing 230, the operator may wish that the additional V0LSUs also dilute the MNC collected during the procedure. In this circumstance, the controller 222 may be arranged to configure the fluid circuit 200 to carry out a preliminary processing cycle 228, as previously described, to collect the additional VOL = us in the PPP container. The controller 222 then configures the fluid circuit 200 to carry out a suspension phase of the MNC 244, to drive the additional VOLSus in the MNC container to achieve the desired dilution of the V0LMNC IV. Process of the Alternative Mononuclear Cell Process Figure 29 shows an alternative modality of a fluid circuit 300, which is prepared for collection and collection of MNCs. Circuit 300 is mostly the same with respect to circuit 200, shown in Figure 6, and common components of the same reference numbers are given. The circuit 300 differs from the circuit 200 in that the second compartment 310 of the container 14 is identical to the compartment 38, and therefore comprises a second blood processing compartment with the same features of the compilation 38. The compartment 310 includes inner seals , as shown for compartment 38 in Figure 4, creating the same blood collection regions for PRP and PRBC, the details of which is not shown in Figure 29. Compartment 310 includes a port 304 for conducting blood in the compartment 310, a port 306 for driving the PRP from the compartment 310, and a port 302 for driving the PRBCs from the compartment 310. The compartment 310 also "includes a tapered ramp 84, as shown in FIGS. 16A and 16B and as described above in relation to compartment 38.

The fluid circuit 300 also differs from the fluid circuit 200 in that the pipes T14, T18, and T19 are not included. In addition, the container of the PRBC is not included. In contrast, the fluid circuit 300 includes several new pipe and clamp paths, as follows: The "path of the pipe T21 is conducted from the PRP output port 306 of the compartment 310 through a new clamp C5 to join the path of the UNC pipe The path of the pipe T22 is conducted from the inlet port of the WB 306 of the compartment 310 through a new air detector D3 and a new clamp C6 to join the path of the pipe T3. trajectory of the pipe T33 is conducted from the outlet port of the PRBC 302 of the compartment 310 through a new clamp C8 to join the path of the pipe T4. The New Clamp C7 is also provided on the T3 pipe upstream of the DI air detector. The New Clamp C9 is also provided in the UNC pipe between the optical detector OS and the junction of the new T21 pipe.

Using circuit 300, controller 222 proceeds through the priming cycle 226 described above, the preliminary processing cycle 228, and the main processing cycle 230 as previously described for circuit 200, through, the accumulation phase. of the MNCs 236. The collection phase of the PRBCs 238 differs by using the circuit 300, in which the PRBCs used to subsequently remove the MNCs from the compartment 38 are processed and collected in the second compartment 310. More particularly, as shown in Figure 30, during the collection phase of the PRBC 238, the controller 222 conducts a whole blood volume of the donor / patient in the second compartment 310. The whole blood volume is extracted by the pump P2 through the tubing TI on the T3 pipe and from there through the "open clamp C6 on the pipe T22, which is led to the compartment 310. The clamp C7 is closed, for blocking uear the conduction of whole blood in compartment 38, where the MNCs have accumulated for collection. The clamp C9 is also closed to block the conduction of the PRP from the compartment 38, thereby maintaining the accumulation of the MNCs motionless in the compartment 38.

In compartment 310, the volume of whole blood is separated in the PRBC and PRP, in the same way that these components are separated in compartment 38. The PRP is led from compartment 310 through the T23 pipe and the clamp open C5 by operating pump P5, to return the donor / patient. The clamp C8 is closed, to retain the PRBCs in the compartment 310. The controller 222 also conducts a different phase of the removal of the MNC 240 using the circuit 300. As shown in Figure 31, during the phase of the removal of the MNCs 240, the controller 222 recirculates a portion of the whole blood drawn back to the donor / patient, while another portion of the whole blood is directed into the compartment 310, following the same path previously described in connection with Figure 30. The controller 222 open clamps C8 and C9, while closing clamp C5. Whole blood entering compartment 310 displaces PRBCs through the outlet port of PRBCs 302 in line T23. The PRBCs of the compartment 310 enter the port of entry of the WB 48 of the compartment 38. As described above, the flow of the PRBCs entering from the outside to the compartment 38 increases the hematocpto of the PRBCs within the compartment 38, causing the MNC accumulated floats to interface 58. As described above, the PRBCs entering from the outside to compartment 38 move the PRP through the port of PRP 46, together with the MNC Region, shown in Figure 31. This Region of the MNC is detected by the optical detector OS and collected in a subsequent process 242, 244, and 246 in the same manner as described for the circuit 200. The various features of the inventions are indicated in the following claims.

It is noted that in relation to this date, the best method known to the applicant to carry out the aforementioned invention, is the conventional one for the manufacture of the objects or products to which it refers. Having described the invention as above, the content of the following is claimed as a priority:

Claims (33)

1. A blood separation system characterized in that it comprises a chamber for rotating approximately on a rotating axis, the chamber includes an entrance region where the whole blood enters for the separation of the condensed red blood cells from a blood sample. constitutes plasma, and of an interface that transports mononuclear cells between the condensed red blood cells of the blood and the plasma constituent, a controller operable in a first mode to conduct whole blood in the region of entry while the condensed red blood cells and the Plasma constituent of the chamber and while maintaining the interface within the chamber, the controller which is also operable in a second mode to remove. the camera interface by conducting the condensed red blood cells in the input region, and an output path to drive the interface removed from the chamber, including a first detector element to locate mononuclear cells at the removed interface and provide a first detection output to locate monenuclear cells. detection output to locate monenuclear cells "

2. A system according to claim 1, characterized in that the output path includes a container in a flow direction downstream of the first detector element, and and wherein the controller directs the collection of the mononuclear cells in the base container, in at least a part, of the first detection output.

3. A system according to claim 1, characterized in that the controller is operable, during the first mode, to remove the plasma constituent from the chamber through the output path.

4. A system according to claim 1, characterized in that, during the second mode, the controller completes the conduction of whole blood in the entrance region.

5. A system according to claim 1, characterized in that, during the second mode, the controller completes the deposition of the condensed red blood cells of the chamber blood.

6. A system according to claim 1, characterized in that the controller circulates the condensed red blood cells withdrawn from the chamber during the first mode to conduct them in the input region during the second mode.

7. A system according to claim 1, characterized in that it additionally includes a reservoir, and where the controller circulates the condensed red blood cells of the reservoir blood to conduct them in the region of entry during the second mode.

8. A system according to claim 7, characterized in that the controller conducts in the reservoir the condensed red blood cells withdrawn from the chamber during the first mode.

9. A system according to claim XI, characterized in that the reservoir comprises a second chamber rotating on an axis in which the red blood cells condensed from the blood are separated from the whole blood centrifugally.

10. A system according to claim 1, characterized in that the first optically detecting element locates mononuclear cells at the removed interface.

11. A system according to claim 1, characterized in that the controller is operable to circulate again the condensed red blood cells withdrawn. to the input _ region during the first mode.

12. A system according to claim 1, characterized in that the controller is operable to again circulate the plasma constituent withdrawn to the input region during the first mode.

13. A system according to claim 1, characterized in that the controller interfaces in the chamber and provides a second detection output.

1.4. A system according to claim 13, characterized in that the second detector element optically locates the interface in the camera.

15. A system according to claim 13, characterized in that the controller is operable, during the first mode, to maintain the interface "in a fixed location in the camera with base, in at least part, of the second detection output.

16. A system according to claim 1, characterized in that the output path includes, in a flow direction downstream of the first detector element, a first branch, a second branch, and a valve element operable in response to the first output. of detection to direct the flow of the first detector element to a selected one of the first branch and the second branch.

17. A system according to claim 16, characterized in that at least one of the first and second branches includes a container.

18. A system according to claim 17, characterized in that, during the second mode, the controller directs the collection of mononuclear cells in the container.

19. A method for the collection of mononuclear cells from whole blood characterized in that it comprises the steps of - (i) rotating a camera on a rotating shaft (ii) conducting the whole blood in an inlet region of the chamber for the separation of the globules condensed reds of the blood, of the plasma constituent, and of the interface which carries the mononuclear cells between the condensed red blood cells of the blood and the plasma constituent, (iii) during step (ii), the condensed red blood cells are removed of the blood and plasma constituent of the chamber while also maintaining the interface within the chamber, (iv) after a prescribed processing period, the condensed red blood cells are driven - in the region of entry to remove the interface of the camera through a path, of output, and (v) locating the mononuclear cells at the removed interface within the output path.

20. A method according to claim 19, characterized in that it additionally includes step (vi) of directing the collection of mononuclear cells in a container communicating with the output path.

21. A method according to claim 19, characterized in that during step (iii), the plasma constituent is removed from the chamber through the exit path.

22. A method according to claim 19, characterized in that during step (iv), the conduction of whole blood in the input region is completed.

23. A method according to claim 19, characterized in that during step (iv), the removal of the condensed red blood cells from the chamber is completed.

24. A method according to claim 19, characterized in that the condensed red blood cells withdrawn from the chamber during step (iii) are conducted in the inlet region during step (iv).

25. A method according to claim 19, characterized in that during step (iv) the condensed red blood cells are circulated from a reservoir to be conducted in the inlet region.

26. A "method according to claim 25, characterized in that at least some of the condensed red blood cells removed from the chamber during step ^ (iii) are transported in the reservoir for circulation to the region of entry during the stage (iv).

27. A method according to claim 25, characterized in that during step (iv), the reservoir rotates while the whole blood is conducted in the reservoir of condensed red blood cells to obtain by centrifugal separation the condensed red blood cells of the blood, which are conducted to the region of entry.

28. A method according to claim 19, characterized in that during the stage (v), mononuclear cells are optically located at the removed interface.

29. A method according to claim 19, characterized in that during the stage (ii), the condensed red blood cells removed during step (iii) are recirculated back to the inlet region.

30. A method according to claim 19, characterized in that during step (i), the plasma constituents removed during step (iii) are recirculated back to the input region.

31. A method according to claim 19, characterized in that during step (iii), the location of the interface in the camera is detected.

32. A method according to claim 31, characterized in that during step (iii), the location of the interface in the camera is detected optically.

33. A method according to claim 31, characterized in that during step (ii), the interface is maintained at a fixed location in the camera based on the detection of the interface.