WO2018021022A1 - Cell treatment device - Google Patents

Abstract

The present disclosure provides a cell treatment device capable of lowering the risk of contamination due to the intermixture of cells before and after a division treatment. The cell treatment device includes: a concentration unit which is provided on the downstream side of a cell supply unit, is connected to the cell supply unit via a first flow passage and performs a concentration treatment for removing the liquid from a cell suspension including cells and a liquid accompanying the cells; and a division unit which is provided on the downstream side of the concentration unit, is connected to the concentration unit via a second flow passage and which performs a division treatment for dividing an aggregate of the cells.

Description

Cell processing equipment

The technology of the present disclosure relates to a cell processing apparatus.

For example, the following techniques are known as techniques relating to cell processing apparatuses (cell culture apparatuses). For example, Japanese Patent Application Laid-Open No. 2010-011792 discloses a culture apparatus, a separation apparatus having filter means connected to the culture tank, and a liquid medium connected to the separation apparatus and stored in the surface of the filter means of the separation apparatus. The culture apparatus provided with the culture-medium transfer tank supplied in the direction parallel to is described. This culture apparatus can reciprocate a liquid medium between a medium transfer tank and a culture tank.

On the other hand, Japanese Patent Application Laid-Open No. 01-206988 discloses a culture system in which a culture medium containing cells or the like is circulated or stored to proliferate cells, and the liquid medium in the culture system can be continuously replaced with a fresh medium. A culture apparatus with a liquid-replenishment / separation means is described. This culturing apparatus is provided with a bypass channel that can temporarily bypass the culture solution to the outside of the culture system. The bypass channel allows the cells in the culture solution to permeate but the cell mass or the like. There is provided a cell lump removing unit made of a cylindrical body with a built-in filter.

In the culture of pluripotent stem cells, when the size of cell aggregates (spheres) generated by culturing cells becomes excessive, the cell aggregates adhere to each other and cells start to differentiate, The cells inside are necrotic. Therefore, in order to prevent the size of the cell aggregate from becoming excessive, a dividing process for dividing the cell aggregate is required at an appropriate time during the cell culture period. In cell culture, the medium is altered by metabolites secreted from the cells. Therefore, it is necessary to perform a medium exchange process in which a used medium in the culture container is replaced with a fresh medium at an appropriate time during the culture period. Further, in cell culture, a concentration process for concentrating the concentration of cells contained in the cell suspension to a desired concentration is required.

The following configuration can be considered as a configuration of a cell processing apparatus that automates a series of cell culture processes including the above-described division processing, medium replacement processing, and concentration processing. For example, the structure which arrange | positions suitably the processing unit and cell culture container for implementing each said process on the cyclic | annular circulation flow path or the branched flow path branched from the circulation flow path can be considered. However, according to this configuration, an annular circulation channel is shared in each process, and it is considered that there is a high risk of so-called contamination in which cells before and after the division process are mixed. For example, some cells attached to the wall surface of the circulation channel while the cell suspension is being transferred through the circulation channel are not subjected to the division process, and these cells are not transmitted through the circulation channel. There is a risk of mixing into cells that have been split. In particular, since cultured cells used for regenerative medicine need to guarantee the homogeneity of cells in the same lot, it is important to reduce the risk of contamination in which cells before and after the division process are mixed.

The technology of the present disclosure provides a cell processing apparatus that can reduce the risk of contamination in which cells before and after division processing are mixed.

A first aspect of the present disclosure is a cell processing apparatus, a cell supply unit that supplies cells, and a downstream side of the cell supply unit, and is connected to the cell supply unit via a first flow path, A concentration unit that performs a concentration process for removing the liquid from the cell suspension containing the cells and the liquid accompanying the cell, and is connected to the concentration unit via the second channel provided downstream of the concentration unit, A dividing unit that performs a dividing process of dividing the aggregate.

In a second aspect of the present disclosure, in the above aspect, the first flow path is a one-way flow path in which the cell suspension passing through the first flow path flows from the upstream side toward the downstream side. Preferably, the second channel is a one-way channel in which the cell suspension passing through the second channel flows from the upstream side toward the downstream side.

According to a third aspect of the present disclosure, in the second aspect, the concentrating unit has one flow port connected to the first flow channel, the other flow port connected to the second flow channel, Concentrate the cell suspension flowing from the flow port or the other flow port, and concentrate from one of the flow ports and the other flow port that is different from the flow port into which the cell suspension flows. A filter unit for discharging the treated cell suspension may be included.

In the fourth aspect of the present disclosure, in the third aspect, the dividing unit may include a discharge port for discharging the cells subjected to the dividing process. In this case, the fourth aspect of the present disclosure may include a third flow path connected to the discharge port and a cell recovery container connected to the third flow path.

The fifth aspect of the present disclosure may include the first bypass flow path having one end connected to the second flow path and the other end connected to the third flow path in the fourth aspect. Good.

According to a sixth aspect of the present disclosure, in the first aspect, the concentration unit includes a first storage container connected to the first flow path, and a second storage container connected to the second flow path. And may be included.

According to a seventh aspect of the present disclosure, in the sixth aspect, the first storage container causes the cell suspension to flow in and out via the first circulation port connected to the first flow path. The second storage container preferably allows the cell suspension to flow in and out through the second flow port connected to the second flow path. Further, according to an eighth aspect of the present disclosure, in the seventh aspect, the first circulation port is provided at a bottom portion of the first storage container, and the second circulation port is a bottom portion of the second storage container. Is preferably provided.

According to a ninth aspect of the present disclosure, in the sixth to eighth aspects, the first pressure control unit that controls the pressure inside the first storage container and the pressure inside the second storage container are controlled. And a second pressure control unit.

According to a tenth aspect of the present disclosure, in the first aspect, one end is connected to the first flow path, the other end is connected to the second flow path, and one end or the other is connected. The cell suspension flowing in from the end of the cell was subjected to stirring treatment, and the stirring treatment was performed from one end and the other end different from the end into which the cell suspension flowed An agitation part for discharging the cell suspension may be included. According to an eleventh aspect of the present disclosure, in the tenth aspect, the stirring unit may include a static mixer.

According to a twelfth aspect of the present disclosure, in the third to sixth and tenth aspects, the second bypass flow having one end connected to the first flow path and the other end connected to the second flow path. It may contain roads.

In a thirteenth aspect of the present disclosure, in the above aspect, the concentration unit may include a plurality of filter units arranged in series or in parallel with each other between the first channel and the second channel. Good.

In the fourteenth and fifteenth aspects of the present disclosure, in the above aspect, the filter unit may perform filtration using a tangential flow method, or may perform separation using centrifugal force.

According to a sixteenth aspect of the present disclosure, in the thirteenth aspect, when the concentration unit includes a plurality of filter units, the concentration unit uses a first filter unit that performs filtration by a tangential flow method and a centrifugal force. And a second filter unit that performs the separation.

The seventeenth aspect of the present disclosure may include a first medium supply unit connected to the first flow path in the above aspect.

The eighteenth aspect of the present disclosure may include a second culture medium supply unit connected to the third flow path in the fourth and fifth aspects.

The nineteenth aspect of the present disclosure may include a cleaning liquid supply unit that supplies a cleaning liquid to the cell supply unit in the above aspect.

According to the above aspect of the present disclosure, it is possible to provide a cell treatment apparatus that can reduce the risk of contamination in which cells before and after the division treatment are mixed.

It is a figure which shows the structure of the cell processing apparatus which concerns on exemplary embodiment of this invention. It is a figure which shows an example of the aspect of filtration in the filter part which concerns on exemplary embodiment of this invention. It is a figure which shows an example of the usage condition of the cell processing apparatus which concerns on exemplary embodiment of this invention. It is a figure which shows the flow of a cell, a culture medium, etc. in case the cell processing apparatus which concerns on exemplary embodiment of this invention performs a division | segmentation process. It is a figure which shows the flow of a cell, a culture medium, etc. in case the cell processing apparatus which concerns on exemplary embodiment of this invention implements a culture medium exchange process. It is a figure which shows the flow of a cell, a culture medium, etc. in case the cell processing apparatus which concerns on exemplary embodiment of this invention implements a concentration process. It is a figure which shows the partial structure of the cell processing apparatus which concerns on other exemplary embodiment of this invention. It is a figure which shows the partial structure of the cell processing apparatus which concerns on other exemplary embodiment of this invention. It is a figure which shows the structure of the cell processing apparatus which concerns on other exemplary embodiment of this invention.

Hereinafter, an exemplary embodiment of the present invention will be described with reference to the drawings. In the drawings, the same or equivalent components and parts are denoted by the same reference numerals, and redundant description is omitted.

[First exemplary embodiment]
FIG. 1 is a diagram showing a configuration of a cell processing apparatus 1 according to a first exemplary embodiment of the present invention. The cell processing apparatus 1 includes a cell supply unit 10, a concentration unit 20, and a division unit 30 as main components. The cell treatment apparatus 1 further includes medium supply units 51 and 52, a cleaning liquid supply unit 60, and a stirring unit 40.

The cell supply unit 10 supplies cells to be processed by the cell processing apparatus 1. The cell supply part 10 is comprised including the storage container which accommodates a cell. The cell supply unit 10 discharges cells from the circulation port 10a provided at the bottom. The circulation port 10a is connected to the first flow path F1 through the valve V1. The valve V1 is opened when the cells accommodated in the cell supply unit 10 are allowed to flow out to the first flow path F1, and is closed otherwise.

The concentration unit 20 is provided on the downstream side of the cell supply unit 10 and is connected to the cell supply unit 10 via the first flow path F1. The concentration unit 20 removes debris such as dead cells that have separated from the liquid and cell aggregates into a single cell from a cell suspension containing cells and a liquid (for example, a medium) associated with the cell, thereby suspending the cell suspension. Concentration treatment is performed to increase the concentration of cells contained in the solution. The concentrating unit 20 includes a filter unit 21, a filtrate recovery container 22, and two storage containers 23A and 23B.

The filter unit 21 has a flow port 21a connected to the first flow path F1 via the valve V2, and a flow port 21b connected to the second flow path F2 via the valve V3. The filter unit 21 performs a concentration process on the cell suspension flowing from the flow port 21a or 21b, and performs a concentration process from a flow port different from the flow port into which the cell suspension flows. Is discharged. For example, the cell suspension that has flowed into the filter unit 21 from the flow port 21a is concentrated in the filter unit 21, and the cell suspension that has been subjected to the concentration process is discharged from the flow port 21b.

The filter unit 21 constitutes, for example, a filtration device that performs filtration by a tangential flow (cross flow) method, and the cell suspension flows along the membrane surface of the filtration membrane disposed inside the filtration device. It is configured as follows. FIG. 2 is a diagram illustrating an example of a filtration mode in the filter unit 21. The filter part 21 has the filtration membrane M which consists of hollow fibers, for example. The filtration membrane M may be a porous membrane, and the material of the filtration membrane M may be a metal, a polymer, a ceramic sintered body, or the like. When a cell suspension containing cells C and a liquid L such as a medium flows into the inside of the filtration membrane M via the flow port 21a or 21b, the liquid L is discharged to the outside of the filtration membrane M and the filtrate collection container 22 Is housed in. On the other hand, the cell C passes through the inside of the filtration membrane M, is discharged from the flow port 21a or 21b, and is collected. The filter unit 21 may perform filtration by a dead end flow method in which the cell suspension flow direction intersects the membrane surface of the filtration membrane. The filter unit 21 may perform separation using centrifugal force.

Valves V2 and V3 are open when the concentration process is performed in the filter unit 21, and are closed otherwise.

The storage containers 23 </ b> A and 23 </ b> B are containers for temporarily storing a cell suspension that is a target of each process performed in the cell processing apparatus 1. The form of the storage containers 23A and 23B is not particularly limited, and for example, a glass or stainless steel container or a container having a plastic bag form can be used.

The storage container 23A has a flow port 23a provided at the bottom thereof, and the cell suspension flows in or out through the flow port 23a. That is, the distribution port 23a serves both as an inlet and an outlet of the storage container 23A. The circulation port 23a is connected to the first flow path F1 through the valve V4. The valve V4 is opened when the cell suspension is allowed to flow into the storage container 23A or when the cell suspension is allowed to flow out of the storage container 23A, and is otherwise closed.

Similarly, the storage container 23B has a flow port 23b provided at the bottom thereof, and the cell suspension flows in or out through the flow port 23b. That is, the circulation port 23b serves both as an inlet and an outlet of the storage container 23B. The circulation port 23b is connected to the second flow path F2 through the valve V5. The valve V5 is opened when the cell suspension is caused to flow into the storage container 23B or when the cell suspension is caused to flow out of the storage container 23B, and is otherwise closed.

The storage container 23A is provided with a pressurizing device 24A and a pressure control device 25A. The pressurizing device 24A has a function of pressurizing the inside of the storage container 23A. The pressure control device 25A controls the pressure at which the pressurizing device 24A pressurizes the inside of the storage container 23A.

Similarly, a pressurizing device 24B and a pressure control device 25B are provided in the storage container 23B. The pressurizing device 24B has a function of pressurizing the inside of the storage container 23B. The pressure control device 25B controls the pressure at which the pressurizing device 24B pressurizes the inside of the storage container 23B.

The cell suspension can be transferred between the storage container 23A and the storage container 23B by pressure control by the pressure control devices 25A and 25B. For example, by making the pressure inside the storage container 23A higher than the pressure inside the storage container 23B, the cell suspension stored in the storage container 23A can be transferred to the storage container 23B. Further, by making the pressure inside the storage container 23B higher than the pressure inside the storage container 23A, the cell suspension stored in the storage container 23B can be transferred to the storage container 23A. In the present embodiment, the case where the cell suspension is transferred by causing a pressure difference in the storage containers 23A and 23B by the pressurizing means that pressurizes the pressure inside the storage containers 23A and 23B is illustrated. It is not limited to the embodiment. As another example, the cell suspension may be transferred by generating a pressure difference in the storage containers 23A and 23B by a decompression unit that reduces the pressure inside the storage containers 23A and 23B. Further, the cell suspension may be transferred by generating a pressure difference in the storage containers 23A and 23B by a combination of the pressurizing means and the decompressing means.

It is also possible to reciprocate the cell suspension between the storage container 23A and the storage container 23B. In this case, the cell suspension passes through the filter unit 21, thereby performing a plurality of concentration processes by the filter unit 21. It becomes possible to carry out continuously over time. Therefore, the concentration of the cell suspension can be adjusted by the number of reciprocating movements of the cell suspension between the storage container 23A and the storage container 23B.

The stirring unit 40 has a function of stirring the flowing cell suspension. One end of the stirring unit 40 is connected to the first flow path F1 through the valve V9, and the other end of the stirring unit 40 is connected to the second flow path F2 through the valve V10. Yes. The agitation unit 40 performs the agitation process on the cell suspension flowing from one end or the other end, and performs the agitation process from an end different from the end into which the cell suspension has flowed. Drain the suspension.

The stirrer 40 preferably has a configuration as a so-called static mixer that does not have a drive unit. For example, the agitator 40 is fixedly installed inside the tubular body and inside the tubular body, and has a spiral flow path inside the tubular body. And a stirring element that forms the structure. The flow path inside the tubular body constituting the static mixer does not necessarily need to be spiral. The static mixer may have a structure in which the inner diameter of the tubular body is changed so that the fluid passing through the inside of the tubular body can be stirred.

Valves V9 and V10 are opened when stirring is performed in the stirring section 40, and are closed otherwise.

The cell treatment device 1 has a bypass channel F20 between the first channel F1 and the second channel F2. That is, one end of the bypass flow path F20 is connected to the first flow path F1 via the valve V11, and the other end of the bypass flow path F20 is connected to the second flow path F2 via the valve V12. ing.

Valves V11 and V12 are opened when the cell suspension is passed through the bypass flow path F20, and are closed otherwise.

As described above, in the cell treatment apparatus 1, the filter unit 21, the stirring unit 40, and the bypass channel F20 are arranged in parallel between the first channel F1 and the second channel F2. Can be selectively used. That is, the cell suspension supplied through the first flow path F1 passes through the filter section 21, the stirring section 40, and the bypass flow path F20 before being transferred to the second flow path F2. Is also possible.

The dividing unit 30 has a function of performing a dividing process of dividing a cell aggregate formed by culturing cells. The dividing process performed in the dividing unit 30 may be a mechanical dividing process or an enzyme process using a cell dissociating enzyme. When the mechanical division process is applied, the dividing unit 30 includes a processing container and a mesh filter provided inside the processing container. By passing the cell suspension containing the cell aggregate through the mesh filter, the cell aggregate is divided into sizes according to the mesh size of the mesh filter. In addition to the mesh filter, a porous plate or a through-hole plate can be used as a means for dividing the cell aggregate. On the other hand, when the division process by the enzyme process is applied, the division unit 30 includes a treatment container in which a cell dissociation enzyme such as trypsin-EDTA (ethylenediaminetetraacetic acid) is accommodated. Cell aggregates are divided by immersing the cell aggregates in a cell dissociation enzyme for a certain period of time

The dividing unit 30 has an inlet 30a connected to the second flow path F2 via the valve V13 and an outlet 30b connected to the third flow path F3 via the valve V14. The dividing unit 30 performs a dividing process on the cell aggregate included in the cell suspension flowing in from the inflow port 30a, and discharges the cell suspension including the cell aggregate subjected to the dividing process from the outflow port 30b. To do.

Valves V13 and V14 are opened when the dividing unit 30 performs division processing, and are otherwise closed.

The cell treatment device 1 has a bypass channel F10 between the second channel F2 and the third channel F3. That is, one end of the bypass flow path F10 is connected to the second flow path F2 via the valve V15, and the other end of the bypass flow path F10 is connected to the third flow path F3 via the valve V16. ing.

Valves V15 and V16 are opened when the cell suspension is passed through the bypass flow path F10, and are closed otherwise.

The cell treatment apparatus 1 has two medium supply parts 51 and 52. The culture medium supply unit 51 includes a storage container that stores a fresh culture medium. The culture medium supply part 51 discharges | emits a culture medium from the discharge port 51a provided in the bottom part. The discharge port 51a is connected to the first flow path F1 through the valve V17. The valve V <b> 17 is opened when the medium stored in the medium supply unit 51 flows out to the first flow path F <b> 1, and is closed otherwise. The discharge port 51a can be disposed at a position other than the bottom of the medium supply unit 51.

The medium supply unit 52 discharges the medium from the discharge port 52a provided at the bottom. The discharge port 52a is connected to the third flow path F3 through the valve V18. The valve V18 is opened when the medium accommodated in the medium supply unit 52 is caused to flow out to the third flow path F3, and is closed otherwise. The discharge port 52a can be disposed at a position other than the bottom of the medium supply unit 52.

The cleaning liquid supply unit 60 includes a storage container that stores a cleaning liquid for cleaning the cell supply unit 10 and the filter unit 21. The cleaning liquid supply unit 60 is connected to the cell supply unit 10 via the valve V19.

The valve V19 is opened when cleaning with the cleaning liquid stored in the cleaning liquid supply unit 60, and is closed otherwise. By opening the valve V19, the cleaning liquid stored in the cleaning liquid supply unit 60 flows into the cell supply unit 10, and the inside of the container constituting the cell supply unit 10 is cleaned. As a result, the cells remaining in the cell supply unit 10 can be scraped and collected by the flow of the washing liquid.

It is also possible to clean the filter part 21 by supplying the cleaning liquid stored in the cleaning liquid supply part 60 to the filter part 21. By flowing the washing liquid through the filter unit 21, cells remaining on the membrane surface of the filtration membrane provided in the filter unit 21 can be scraped and collected by the washing liquid flow. In addition, it is possible to use the same culture medium as the culture medium accommodated in the culture medium supply units 51 and 52 as the cleaning liquid.

The cell collection container 100 is a container for collecting the cell suspension processed by the cell processing apparatus 1. The form of the cell collection container 100 is not particularly limited, and for example, a glass or stainless steel container or a container having a plastic bag form can be used. As shown in FIG. 1, by connecting the cell collection container 100 to the third flow path F3, it is possible to collect the cells treated in the cell treatment apparatus 1 in the cell collection container 100. The cell collection container 100 has a flow port 100a provided at the bottom thereof, and the cell suspension processed by the cell processing apparatus 1 flows through the flow port 100a. When the cell suspension treated in the cell treatment apparatus 1 is collected in the cell collection container 100, the flow port 100a of the cell collection container 100 is connected to the third flow path F3 via the valve V22. The valve V22 is opened when the treated cell suspension is allowed to flow into the cell collection container 100, and is closed otherwise.

In addition, when the cell suspension accommodated in the cell collection container 100 is processed by the cell processing apparatus 1, the cell collection container 100 is connected to the cell supply unit 10 as shown in FIG. In this case, the valve V22 is opened when the cell suspension accommodated in the cell collection container 100 is allowed to flow into the cell supply unit 10, and is closed otherwise.

In the cell treatment apparatus 1, it is preferable that the connection part X between the pipes constituting each flow path is constituted by a sterilization connector.

In the cell treatment apparatus 1, the flow direction of the cell suspension passing through the first flow path F1 is controlled so as to always be the direction from the cell supply unit 10 toward the concentration unit 20 (the direction of the arrow shown in FIG. 1). It is preferred that

The liquid flow of the cell suspension passing through the first channel F1 may be formed by, for example, a pump (not shown) disposed on the first channel F1. In this case, it is possible to control the flow direction of the cell suspension passing through the first flow path F1 by the rotation direction of the pump arranged on the first flow path F1.

Further, the liquid flow of the cell suspension passing through the first flow path F1 is formed by a differential pressure between the pressure inside the cell supply unit 10 or the medium supply unit 51 and the pressure inside the storage container 23A or 23B. May be. In this case, the same mechanism as the pressurization devices 24A and 24B and the pressure control devices 25A and 25B provided in the storage containers 23A and 23B is provided in the cell supply unit 10 and the culture medium supply unit 51. For example, by making the pressure inside the cell supply unit 10 higher than the pressure inside the storage container 23A or 23B, the flow direction of the cell suspension passing through the first flow path F1 is changed to the cell supply unit 10. The direction toward the concentrating unit 20 can be made. The means for forming the liquid flow of the cell suspension passing through the first flow path F1 is not limited to the application of pressure to the inside of the cell supply unit 10, the medium supply unit 51, and the storage containers 23A and 23B. Pressure may be applied from the outside.

Note that a valve V20 may be provided at the downstream end of the first flow path F1 in order to prevent the back flow of the cell suspension in the first flow path F1. The valve V20 is opened when the cell suspension is passed through the first flow path F1, and is closed otherwise. Note that a check valve, a check valve or the like can be used as another means for preventing the back flow of the cell suspension in the first flow path F1. By using a check valve or a check valve, the liquid flow from the upstream side to the downstream side can be passed through the first flow path F1, while the liquid flow in the reverse direction can be blocked.

In the cell treatment device 1, the flow direction of the cell suspension passing through the second flow path F2 is always the direction from the concentration unit 20, the stirring unit 40, or the bypass flow channel F20 toward the dividing unit 30 or the bypass flow channel F10 ( It is preferably controlled so that the direction of the arrow shown in FIG.

The liquid flow of the cell suspension passing through the second flow path F2 may be formed by, for example, a pump (not shown) disposed on the second flow path F2. In this case, it is possible to control the flow direction of the cell suspension passing through the second flow path F2 by the rotation direction of the pump disposed on the second flow path F2.

Further, the liquid flow of the cell suspension passing through the second flow path F2 may be formed by a differential pressure between the pressure inside the storage container 23A or 23B and the pressure inside the cell collection container 100. Or you may form by the differential pressure | voltage of the pressure inside the storage container 23A or 23B, and the pressure inside the division part 30. FIG. In this case, the cell recovery container 100 or the dividing unit 30 is provided with the same mechanism as the pressurization devices 24A and 24B and the pressure control devices 25A and 25B provided in the storage containers 23A and 23B. By making the pressure inside the storage container 23A or 23B higher than the pressure inside the cell collection container 100 or the dividing unit 30, the flow direction of the cell suspension passing through the second flow path F2 is changed to the concentration unit. 20, It can be set as the direction which goes to the division | segmentation part 30 or the bypass flow path F10 from the stirring part 40 or the bypass flow path F20.

In addition, in order to prevent the back flow of the cell suspension in the second flow path F2, a valve V21 may be provided at the upstream end of the second flow path F2. The valve V21 is opened when the cell suspension is passed through the second flow path F2, and is closed otherwise. Note that a check valve or a check valve can be used as another means for preventing the back flow of the cell suspension in the second flow path F2. By using a check valve or a check valve, the liquid flow from the upstream side to the downstream side can be passed through the second flow path F2, while the liquid flow in the reverse direction can be blocked.

In the cell treatment device 1, the flow direction of the cell suspension passing through the third flow path F3 is always the direction from the dividing section 30 or the bypass flow path F10 toward the cell collection container 100 (indicated by the arrow shown in FIG. 1). Direction).

The liquid flow of the cell suspension passing through the third flow path F3 may be formed by, for example, a pump (not shown) arranged on the third flow path F3. In this case, it is possible to control the flow direction of the cell suspension passing through the third flow path F3 by the rotation direction of the pump disposed on the third flow path F3.

Further, the liquid flow of the cell suspension passing through the third flow path F3 may be formed by a differential pressure between the pressure inside the storage container 23A or 23B and the pressure inside the cell collection container 100. In this case, the cell recovery container 100 is provided with the same mechanism as the pressurization devices 24A and 24B and the pressure control devices 25A and 25B provided in the storage containers 23A and 23B. For example, by making the pressure inside the storage container 23A or 23B higher than the pressure inside the cell collection container 100, the flow direction of the cell suspension passing through the third flow path F3 is changed to the dividing unit 30 or the bypass. The direction can be the direction from the flow path F10 toward the cell collection container 100.

The control unit 70 performs opening / closing control of the valves V1 to V21. When the liquid flow of the cell suspension is formed by a pump (not shown) arranged on the first flow path F1, the second flow path F2, and the third flow path F3, the control unit 70 The operation control of each pump is performed in conjunction with the opening / closing operation of the valves V1 to V21. Further, when the liquid flow of the cell suspension is formed by the differential pressure among the cell supply unit 10, the storage containers 23A and 23B, the dividing unit 30, and the cell collection container 100, the pressure adjustment mechanism provided in each of these units Is controlled in conjunction with the opening and closing operations of the valves V1 to V21. That is, the control unit 70 performs liquid feeding control so that the cell suspension flows through a predetermined path in each process performed in the cell processing apparatus 1. Note that the control of the opening and closing of the valve V22 associated with the cell collection container 100 may be performed by the control unit 70 or manually.

Hereinafter, an example of processing performed in the cell processing apparatus 1 according to the exemplary embodiment will be described. The cell treatment apparatus 1 can perform, for example, a division process, a medium exchange process, and a concentration process exemplified below.

<Division processing>
In the culture of pluripotent stem cells, when the size of cell aggregates called spheres generated by culturing cells becomes excessive, cell aggregates adhere to each other and cells start to differentiate, Problems arise such as necrosis of the cells inside. Therefore, in order to prevent the size of the cell aggregate from becoming excessive, a dividing process for dividing the cell aggregate is required at an appropriate time during the culture period. The cell processing apparatus 1 performs the above-described division processing as follows. In the following description, a case where the dividing process in the dividing unit 30 is a mechanical dividing process will be exemplified. FIG. 4 is a diagram illustrating a flow of cells, a medium, and the like when the cell processing apparatus 1 performs a division process. FIG. 4 shows the correspondence between the flow of cells and culture medium, etc., and the following processing steps.

When carrying out the division process in the cell treatment apparatus 1, the cell aggregates to be subjected to the division process are accommodated in the cell supply unit 10 and supplied from the cell supply unit 10.

In step S1, the valves V1, V20, V11, V12, and V5 are opened, and are accommodated in the cell supply unit 10 by controlling the operation of the pump or controlling the differential pressure between the cell supply unit 10 and the storage container 23B. The cell suspension is transferred to the storage container 23B via the first flow path F1 and the bypass flow path F20.

In step S2, the valve V19 is opened. As a result, the cleaning liquid stored in the cleaning liquid supply unit 60 flows into the cell supply unit 10 and the cell supply unit 10 is cleaned. Thereby, the cells (cell aggregates) remaining in the cell supply unit 10 are scraped off by the flow of the cleaning liquid.

In step S3, the valves V1, V20, V11, V12 and V5 are opened and remain in the cell supply unit 10 by controlling the operation of the pump or controlling the differential pressure between the cell supply unit 10 and the storage container 23B. The cells (cell aggregates) and the washing liquid are transferred to the storage container 23B via the first flow path F1 and the bypass flow path F20. That is, the cells (cell aggregates) remaining in the cell supply unit 10 are stored in the storage container 23B together with the cells (cell aggregates) transferred to the storage container 23B in step S1.

In step S4, the valves V5, V3, V2, and V4 are opened, and the cells (cell aggregates) stored in the storage container 23B, the washing liquid, and the control are performed by controlling the differential pressure between the storage container 23A and the storage container 23B. The cell suspension containing the medium is transferred to the storage container 23A via the filter unit 21. The cell suspension is concentrated while passing through the filter unit 21.

The filter unit 21 performs a concentration process for removing the washing solution and the medium from the cell suspension containing the cells (cell aggregates), the washing solution and the medium flowing in from the flow port 21b to increase the concentration of the cells. The washing liquid and the culture medium are collected in the filtrate collection container 22, and the concentrated cell suspension is discharged from the circulation port 21a and stored in the storage container 23A. In addition, you may implement the concentration process by the filter part 21 continuously in multiple times by reciprocating a cell suspension between the storage container 23A and the storage container 23B.

In step S5, the valves V19, V1, V20, V11, V12, and V5 are opened and accommodated in the cleaning liquid supply unit 60 by controlling the operation of the pump or controlling the differential pressure between the cell supply unit 10 and the storage container 23B. The washed liquid thus transferred is transferred to the storage container 23B via the cell supply unit 10, the first flow path F1, and the bypass flow path F20. Subsequently, the valves V5, V3, V2, and V4 are opened, and the cleaning liquid stored in the storage container 23B flows to the filter unit 21 by the differential pressure control between the storage container 23A and the storage container 23B. As the washing liquid flows into the filter unit 21, cells (cell aggregates) remaining on the filtration membrane of the filter unit 21 are scraped by the washing liquid flow, and the cells (cell coagulation) transferred to the storage container 23A in step S4. Is collected in the storage container 23A.

In step S6, the valves V17, V20, and V4 are opened, and the fresh medium stored in the medium supply unit 51 is controlled by the pump operation control or the differential pressure control between the medium supply unit 51 and the storage container 23A. It is transferred to the storage container 23A via the first flow path F1. Thereby, a fresh culture medium is supplied to the cells (cell aggregates) stored in the storage container 23A.

In step S7, the valves V4, V9, V10, V21, V13, V14, and V22 are opened and stored in the storage container 23A by pump operation control or differential pressure control between the storage container 23A and the cell collection container 100. The cell suspension thus obtained is transferred to the cell collection container 100 via the stirring unit 40, the second channel F2, the dividing unit 30, and the third channel F3.

The cell suspension is stirred by passing through the stirring unit 40. Thereby, cells (cell aggregates) are uniformly dispersed in the medium.

The cell suspension that has passed through the stirring unit 40 flows into the processing container of the dividing unit 30 from the inflow port 30a. The cells (cell aggregates) contained in the suspension flow into the processing container of the dividing unit 30 and pass through the mesh filter provided in the processing container, thereby being divided into sizes according to the mesh size of the mesh filter. . The cell suspension containing the cells (cell aggregates) subjected to the division treatment is discharged from the outflow port 30b and is stored in the cell collection container 100 via the third flow path F3.

In step S8, the valves V18 and V22 are opened, and the fresh medium stored in the medium supply unit 52 is controlled by the pump operation control or the differential pressure control between the medium supply unit 52 and the cell collection container 100. It is transferred to the cell collection container 100 via the third flow path F3. Thereby, the quantity of the culture medium in the cell collection container 100 is adjusted. If adjustment of the amount of medium is not required, the medium supply from the medium supply unit 52 may be omitted.

After the division process is completed, the cells collected in the cell collection container 100 are transferred to a cell culture container (not shown), and the cell culture is continued. Further, all or a part of the cell suspension containing the cells (cell aggregates) subjected to the dividing process by the dividing unit 30 may be stored in a storage container and stored frozen.

<Medium medium replacement process>
In cell culture, the medium is altered by metabolites secreted from the cells. For this reason, a medium replacement process is required to replace a used medium with a fresh medium at an appropriate time during the culture period. The cell treatment apparatus 1 performs the above-described medium exchange process as follows. FIG. 5 is a diagram illustrating a flow of cells, a medium, and the like when the cell processing apparatus 1 performs a medium replacement process. FIG. 5 shows the correspondence between the flow of cells and culture medium, etc., and the following processing steps.

When replacing a used medium stored in the cell collection container 100 with a fresh medium, the cell collection container 100 is connected to the cell supply unit 10 as shown in FIG. Thereafter, by opening the valve V22, the cell suspension containing the used medium contained in the cell collection container 100 is put into the cell supply unit 10. Thereafter, the cell collection container 100 is connected to the third flow path F3 as shown in FIG.

In step S11, the valves V1, V20, V11, V12, and V5 are opened, and are accommodated in the cell supply unit 10 by pump operation control or differential pressure control between the cell supply unit 10 and the storage container 23B. The cell suspension containing the used medium is transferred to the storage container 23B via the first channel F1 and the bypass channel F20.

In step S12, the valve V19 is opened. As a result, the cleaning liquid stored in the cleaning liquid supply unit 60 flows into the cell supply unit 10 and the cell supply unit 10 is cleaned. Thereby, the cells remaining in the cell supply unit 10 are scraped off by the flow of the cleaning liquid.

In step S13, the valves V1, V20, V11, V12, and V5 are opened and remain in the cell supply unit 10 by controlling the operation of the pump or controlling the differential pressure between the cell supply unit 10 and the storage container 23B. The cells and the washing liquid are transferred to the storage container 23B via the first flow path F1 and the bypass flow path F20. That is, the cells remaining in the cell supply unit 10 are stored in the storage container 23B together with the cells transferred to the storage container 23B in step S11.

In step S14, the valves V5, V3, V2, and V4 are opened, and the cells stored in the storage container 23B, the washing solution, and the used medium are included by controlling the differential pressure between the storage container 23A and the storage container 23B. The cell suspension is transferred to the storage container 23A via the filter unit 21. The cell suspension is concentrated while passing through the filter unit 21.

The filter unit 21 performs a concentration process to remove the washing solution and the used medium from the cell suspension containing the cells, the washing solution, and the used medium that have flowed from the flow port 21b, thereby increasing the cell concentration. The washing liquid and the used culture medium are collected in the filtrate collection container 22, and the concentrated cell suspension is discharged from the circulation port 21a and stored in the storage container 23A. In addition, you may implement the concentration process by the filter part 21 continuously in multiple times by reciprocating a cell suspension between the storage container 23A and the storage container 23B.

In step S15, the valves V1, V20, V11, V12, and V5 are opened, and are accommodated in the cleaning liquid supply unit 60 by pump operation control or differential pressure control between the cell supply unit 10 and the storage container 23B. The cleaning solution is transferred to the storage container 23B via the cell supply unit 10, the first flow path F1, and the bypass flow path F20. Subsequently, the valves V5, V3, V2, and V4 are opened, and the cleaning liquid stored in the storage container 23B flows to the filter unit 21 by the differential pressure control between the storage container 23A and the storage container 23B. As the cleaning liquid flows through the filter unit 21, the cells remaining on the filtration membrane of the filter unit 21 are scraped off by the liquid flow of the cleaning liquid and stored in the storage container 23A together with the cells transferred to the storage container 23A in step S14. .

In step S16, the valves V17, V20, and V4 are opened, and the fresh medium stored in the medium supply unit 51 by controlling the operation of the pump or controlling the differential pressure between the medium supply unit 51 and the storage container 23A. Is transferred to the storage container 23A via the first flow path F1. Thereby, a fresh culture medium is supplied to the cells stored in the storage container 23A.

In step S17, the valves V4, V9, V10, V21, V15, V16, and V22 are opened, and stored in the storage container 23A by pump operation control or differential pressure control between the storage container 23A and the cell collection container 100. The cell suspension thus obtained is transferred to the cell collection container 100 via the stirring unit 40, the second flow path F2, the bypass flow path F10, and the third flow path F3.

The cell suspension is stirred by passing through the stirring unit 40. Thereby, the cells are uniformly dispersed in the medium.

The cell suspension that has passed through the stirring unit 40 is accommodated in the cell collection container 100 via the second flow path F2, the bypass flow path F10, and the third flow path F3.

In step S18, the valves V18 and V22 are opened, and the fresh medium contained in the medium supply unit 52 is controlled by controlling the operation of the pump or controlling the differential pressure between the medium supply unit 52 and the cell collection container 100. 3 is transferred to the cell collection container 100 via the flow path F3. Thereby, the quantity of the culture medium in the cell collection container 100 is adjusted. If adjustment of the amount of medium is not required, the medium supply from the medium supply unit 52 may be omitted.

<Concentration treatment>
As described above, the cell suspension concentration process is performed along with the division process and the medium exchange process. However, according to the cell processing apparatus 1 according to the exemplary embodiment, the cell suspension concentration process is performed. Can also be carried out alone. FIG. 6 is a diagram illustrating the flow of cells, culture medium, and the like when the cell processing apparatus 1 performs the concentration process alone. FIG. 6 shows the correspondence between the flow of cells and culture medium and the following processing steps.

When the cell suspension in the cell collection container 100 is concentrated, the cell collection container 100 is connected to the cell supply unit 10 as shown in FIG. Thereafter, by opening the valve V22, the cell suspension containing the cells and the medium contained in the cell collection container 100 is put into the cell supply unit 10. Thereafter, the cell collection container 100 is connected to the third flow path F3 as shown in FIG. In addition, it is also possible to perform a concentration process for objects other than the cell suspension in the cell collection container 100.

In step S31, the valves V1, V20, and V4 are opened, and the cell suspension accommodated in the cell supply unit 10 is controlled by pump operation control or differential pressure control between the cell supply unit 10 and the storage container 23A. Then, it is transferred to the storage container 23A via the first flow path F1.

In step S32, the valve V19 is opened. As a result, the cleaning liquid stored in the cleaning liquid supply unit 60 flows into the cell supply unit 10 and the cell supply unit 10 is cleaned. Thereby, the cells remaining in the cell supply unit 10 are scraped off by the flow of the cleaning liquid.

In step S33, the valves V1, V20 and V4 are opened, and the cells and washing liquid remaining in the cell supply unit 10 by controlling the operation of the pump or controlling the differential pressure between the cell supply unit 10 and the storage container 23A. Is transferred to the storage container 23A via the first flow path F1. That is, the cells remaining in the cell supply unit 10 are stored in the storage container 23A together with the cells transferred to the storage container 23A in step S31.

In step S34, the valves V4, V2, V3, V21, V15, V16, and V22 are opened and stored in the storage container 23A by pump operation control or differential pressure control between the storage container 23A and the cell collection container 100. The cell suspension containing the cells, the washing solution, and the culture medium is transferred into the cell collection container 100 via the filter unit 21, the second channel F2, the bypass channel F10, and the third channel F3. . The cell suspension is concentrated while passing through the filter unit 21.

The filter unit 21 removes debris such as dead cells that have been detached from the washing solution, the medium, and the cell aggregate from the cell suspension containing the cells, the washing solution, and the medium that have flowed in from the flow port 21a, thereby forming a single cell concentration. To increase the concentration. The washing liquid, the medium, and the debris are collected in the filtrate collection container 22, and the concentrated cell suspension is discharged from the circulation port 21b, and the second flow path F2, the bypass flow path F10, and the third flow path F3. To be accommodated in the cell collection container 100. In addition, you may implement the concentration process by the filter part 21 continuously in multiple times by reciprocating a cell suspension between the storage container 23A and the storage container 23B.

In step S35, the valves V19, V1, V20, V2, V3, V21, V15, V16, and V22 are opened, and pump pressure control or differential pressure control between the cell supply unit 10 and the cell collection container 100 is performed. The cleaning liquid stored in the cleaning liquid supply unit 60 is recovered through the cell supply unit 10, the first channel F1, the filter unit 21, the second channel F2, the bypass channel F10, and the third channel F3. It is transferred to the container 100. As the washing liquid flows into the filter unit 21, cells remaining on the filtration membrane of the filter unit 21 are scraped off by the washing liquid flow, and are accommodated in the cell collection container 100 together with the cells contained in the cell collection container 100 in step S34. Is done.

In step S36, the valves V18 and V22 are opened, and the fresh medium contained in the medium supply unit 52 is controlled by the pump operation control or the differential pressure control between the medium supply unit 52 and the cell collection container 100. 3 is transferred to the cell collection container 100 via the flow path F3. Thereby, the quantity of the culture medium in the cell collection container 100 is adjusted. If adjustment of the amount of medium is not required, the medium supply from the medium supply unit 52 may be omitted.

As is clear from the above description, in the cell processing apparatus 1 according to the exemplary embodiment of the present invention, the concentration unit 20 is provided on the downstream side of the cell supply unit 10, and the cells are passed through the first flow path F1. It is connected to the supply unit 10. The dividing unit 30 is provided on the downstream side of the concentration unit 20 and is connected to the concentration unit 20 via the second flow path F2. That is, in the cell treatment apparatus 1, the cell supply unit 10, the concentration unit 20, and the division unit 30 are configured to be connected in series in this order. As shown in FIG. 1, when the cell collection container 100 is connected to the third flow path F3, the cell supply unit 10, the concentration unit 20, the dividing unit 30, and the cell collection container 100 are connected in series in this order. It becomes the composition which was done. Moreover, in the cell treatment apparatus 1, the flow direction of the cell suspension passing through the first flow path F1, the second flow path F2, and the third flow path F3 is only one direction. That is, the first flow path F1, the second flow path F2, and the third flow path F3 are one-way flow paths. Thus, by connecting the cell supply unit 10, the concentration unit 20, and the dividing unit 30 in series via the one-way flow path in the cell processing apparatus 1, the risk of contamination in which cells before and after the dividing process are mixed is reduced. It becomes possible to do.

For example, when the division process is performed, the bypass flow path F10 is blocked, and only the cells that have passed through the division unit 30 are collected in the cell collection container 100. In other words, when the division process is performed, the cells are not collected in the cell collection container 100 without passing through the division unit 30. Therefore, it is possible to prevent the occurrence of contamination in which the cells before the division process and the cells after the division process are mixed.

Moreover, according to the cell processing apparatus 1 which concerns on exemplary embodiment of this invention, before the cell supplied from the cell supply part 10 is collect | recovered by the cell collection container 100, said division | segmentation process and culture medium replacement | exchange are performed. Either one of the treatment and the concentration treatment is performed. That is, the above division process, medium exchange process, and concentration process are performed independently of each other. Therefore, after completion of each treatment, the cell collection container 100 can be detached from the cell treatment apparatus 1 and the treatment units constituting the cell treatment apparatus 1 and the pipes constituting each flow path can be washed. In this way, by configuring the device to divide and implement each treatment, contamination in which cells before and after the division treatment are mixed as compared to a device configuration that continuously performs each treatment using a circulation channel. Risk can be reduced.

In the cell treatment device 1 according to the exemplary embodiment of the present invention, the dividing unit 30 and the bypass channel F10 are arranged in parallel between the second channel F2 and the third channel F3. . Thereby, when performing culture medium exchange processing or concentration processing, the processed cell suspension can be collected in the cell collection container 100 without going through the dividing unit 30.

In the cell treatment device 1 according to the exemplary embodiment of the present invention, the filter unit 21, the stirring unit 40, and the bypass channel F20 are arranged in parallel between the first channel F1 and the second channel F2. Has been placed. The cell treatment device 1 includes a storage container 23A connected to the first flow path F1 and a storage container 23B connected to the second flow path F2, and between the storage container 23A and the storage container 23B. Thus, the cell suspension can be transferred to each other. According to this structure, the filter part 21, the stirring part 40, and the bypass flow path F20 can be used as a bidirectional | two-way flow path, for example, the concentration process in the filter part 21 can be performed continuously several times. It becomes. Further, for example, the concentration process in the filter unit 21 and the stirring process in the stirring unit 40 can be continuously performed. Furthermore, the mutual transfer of the cell suspension between the storage container 23A and the storage container 23B can be performed via the bypass channel F20, and the cell suspension supplied from the cell supply unit 10 It is also possible to transfer to the storage container 23B via the bypass flow path F20.

In addition, in the cell processing apparatus 1 according to the exemplary embodiment of the present invention, the storage container 23A allows the cell suspension to flow in and out through the flow port 23a provided at the bottom thereof. Similarly, the storage container 23B allows the cell suspension to flow in and out through the circulation port 23b provided at the bottom. In this way, the circulation ports 23a and 23b serve as the inflow port and the outflow port, so that one pipe can be connected to the storage containers 23A and 23B. Thereby, compared with the case where storage container 23A and 23B are provided with an inflow port and an outflow port separately, the number of piping can be reduced and the loss of the cell by a cell adhering to piping can be reduced.

Further, by providing the circulation ports 23a and 23b at the bottoms of the storage containers 23A and 23B, respectively, damage to the cells when the cell suspension is allowed to flow into the storage containers 23A and 23B can be suppressed. In other words, by providing the flow ports 23a and 23b at the bottoms of the storage containers 23A and 23B, respectively, the cell suspension is stored in such a manner that the cell suspension is injected into the cell suspension already accumulated at the bottoms of the storage containers 23A and 23B. Cell suspension can be flowed into containers 23A and 23B. Thereby, compared with the case where a cell suspension is made to flow into storage container 23A and 23B in the aspect which drops a cell suspension from the upper part of a storage container, the damage to a cell can be reduced and the loss of a cell is reduced. be able to.

In addition, the cell treatment device 1 according to the exemplary embodiment of the present invention includes a medium supply unit 51 connected to the first flow path F1. Thereby, the culture medium exchange process which replaces a used culture medium with a fresh culture medium is attained. Moreover, the cell processing apparatus 1 which concerns on exemplary embodiment of this invention has the culture medium supply part 52 connected to the 3rd flow path F3. Thereby, a culture medium can be added to the treated cell suspension recovered in the cell recovery container 100, and the amount of the culture medium can be adjusted after each process performed in the cell processing apparatus 1.

In addition, the cell processing apparatus 1 according to the exemplary embodiment of the present invention includes a cleaning liquid supply unit 60 connected to the cell supply unit 10. Thereby, washing | cleaning of the cell supply part 10, the filter part 21, etc. can be performed during a process, and the cells which remain | survive in the cell supply part 10, the filter part 21, etc. can be collect | recovered. Thereby, the loss of cells can be reduced.

In the above-described exemplary embodiment, the mode in which the cell collection container 100 is attached to the cell processing apparatus 1 is exemplified. However, the mode in which the cell collection container 100 is attached to the third flow path F3 is used as the cell processing. It can also be regarded as a form of the device.

[Second exemplary embodiment]
7 and 8 are views showing a partial configuration of the cell processing apparatus according to the second exemplary embodiment of the present invention. The cell treatment apparatus may include a plurality of filter units. For example, as shown in FIG. 7, the filter unit 21A and the filter unit 21B are provided between the first channel F1 and the second channel F2. May be arranged in parallel. According to this configuration, the concentration process can be performed in parallel in the filter units 21A and 21B, and the concentration process can be performed efficiently.

Further, as shown in FIG. 8, the filter part 21A and the filter part 21B may be arranged in series between the first flow path F1 and the second flow path F2. According to this configuration, it is possible to continuously perform the concentration process by the filter unit 21A and the concentration process by the filter unit 21B with a single liquid feeding.

7 and FIG. 8, the filtering methods in the filter units 21A and 21B may be different from each other. For example, the filter unit 21A may be a filtration device that performs filtration by a tangential flow method, and the filter unit 21B may be a filtration device that performs separation using centrifugal force. Further, the filtering methods in the filter units 21A and 21B may be the same as each other.

[Third exemplary embodiment]
Each component of the 1st cell processing apparatus 1 shown in FIG. 1 can be suitably abbreviate | omitted according to a cell culture protocol. FIG. 9 is a diagram showing a configuration of a cell processing apparatus 1A according to the third exemplary embodiment of the present invention, in which the configuration of the cell processing apparatus 1 according to the first exemplary embodiment is simplified.

The cell treatment apparatus 1A corresponds to a configuration in which the agitation unit 40, the cleaning liquid supply unit 60, and the culture medium supply unit 52 are omitted from the cell treatment apparatus 1 according to the first exemplary embodiment. Thus, by simplifying the configuration, it is possible to save the space of the cell processing apparatus. Even in the cell processing apparatus 1A having a simplified configuration, it is possible to perform the same division process, medium replacement process, and concentration process as those of the cell processing apparatus 1 according to the first exemplary embodiment.

The cell processing apparatus 1 and 1A are examples of the cell processing apparatus in the technology of the present disclosure. The cell supply unit 10 is an example of a cell supply unit in the technology of the present disclosure. The concentration unit 20 is an example of a concentration unit in the technology of the present disclosure. The dividing unit 30 is an example of a dividing unit in the technology of the present disclosure. The first flow path F1 is an example of the first flow path in the technology of the present disclosure. The second flow path F2 is an example of a second flow path in the technology of the present disclosure. The third flow path F3 is an example of a third flow path in the technology of the present disclosure. The filter unit 21 is an example of a filter unit in the technology of the present disclosure. The cell collection container 100 is an example of cell collection in the technology of the present disclosure. The bypass channel F10 is an example of a first bypass channel in the technology of the present disclosure. The bypass channel F20 is an example of a second bypass channel in the technology of the present disclosure. The storage container 23A is an example of a first storage container in the technology of the present disclosure. The storage container 23B is an example of a second storage container in the technology of the present disclosure. The distribution port 23a is an example of a first distribution port in the technology of the present disclosure. The distribution port 23b is an example of a second distribution port in the technology of the present disclosure. The stirring unit 40 is an example of a stirring unit in the technology of the present disclosure. Pressurization device 24A and pressure control device 25A are examples of the 1st pressure control part in the art of this indication. The pressurizing device 24B and the pressure control device 25B are an example of a second pressure control unit in the technology of the present disclosure. The culture medium supply unit 51 is an example of a first culture medium supply unit in the technology of the present disclosure. The culture medium supply unit 52 is an example of a second culture medium supply unit in the technology of the present disclosure. The cleaning liquid supply unit 60 is an example of a cleaning liquid supply unit in the technology of the present disclosure.

The entire disclosure of Japanese Application 2016-150671 is incorporated herein by reference.

All documents, patent applications, and technical standards mentioned in this specification are to the same extent as if each individual document, patent application, and technical standard were specifically and individually described to be incorporated by reference, Incorporated herein by reference.

Claims (19)

A cell supply section for supplying cells;
A concentration process that is provided on the downstream side of the cell supply unit and is connected to the cell supply unit via a first flow path, and removes the liquid from the cell suspension containing the cells and the liquid accompanying the cells. A concentration unit for performing
A dividing unit that is provided on the downstream side of the concentrating unit and connected to the concentrating unit via a second flow path, and performs a dividing process of dividing the aggregate of the cells;
A cell processing apparatus comprising: The first flow channel is a one-way flow channel in which a cell suspension passing through the first flow channel flows from the cell supply unit side toward the concentration unit side,
The cell treatment according to claim 1, wherein the second flow path is a one-way flow path in which a cell suspension passing through the second flow path flows from the concentration section side toward the division section side. apparatus. The concentrating unit has one flow port connected to the first flow channel, the other flow port connected to the second flow channel, and cells flowing from the one flow port or the other flow port. The cell suspension that has been subjected to the concentration treatment from a flow port different from the flow port into which the cell suspension has flowed out of the one flow port and the other flow port, by performing the concentration treatment on the suspension The cell processing apparatus according to claim 2, further comprising a filter unit that discharges the liquid. The dividing unit includes a discharge port for discharging cells subjected to the dividing process,
A third flow path connected to the outlet;
A cell collection container connected to the third flow path;
The cell treatment apparatus according to claim 3, further comprising: 5. The cell treatment device according to claim 4, further comprising a first bypass channel having one end connected to the second channel and the other end connected to the third channel. The said concentration part further contains the 1st storage container connected to the said 1st flow path, and the 2nd storage container connected to the said 2nd flow path. The cell treatment apparatus according to any one of the above. One end is connected to the first flow path, the other end is connected to the second flow path, and the cell suspension flows from the one end or the other end. The agitation unit that performs the agitation process and discharges the cell suspension subjected to the agitation process from an end part of the one end part and the other end part different from the end part into which the cell suspension has flowed The cell treatment device according to any one of claims 3 to 6, further comprising: 8. The method according to claim 3, further comprising a second bypass flow path having one end connected to the first flow path and the other end connected to the second flow path. Cell processing equipment. The first storage container allows the cell suspension to flow in and out through the first flow port connected to the first flow path,
The cell processing apparatus according to claim 6, wherein the second storage container allows a cell suspension to flow in and out through a second circulation port connected to the second flow path. The first circulation port is provided at the bottom of the first storage container,
The cell processing apparatus according to claim 9, wherein the second circulation port is provided at a bottom portion of the second storage container. A first pressure control unit for controlling the pressure inside the first storage container;
A second pressure control unit for controlling the pressure inside the second storage container;
The cell treatment apparatus according to any one of claims 6, 9, and 10, further comprising: The said concentration part contains the several filter part containing the said filter part arrange | positioned mutually in series or in parallel between the said 1st flow path and the said 2nd flow path. The cell treatment apparatus according to any one of the above. The cell processing apparatus according to any one of claims 3 to 12, wherein the filter unit performs filtration by a tangential flow method. The cell processing apparatus according to any one of claims 3 to 12, wherein the filter unit performs separation using centrifugal force. The cell processing apparatus according to claim 12, wherein the plurality of filter units include a first filter unit that performs filtration by a tangential flow method and a second filter unit that performs separation using centrifugal force. The cell processing apparatus according to claim 7, wherein the stirring unit includes a static mixer. The cell treatment apparatus according to any one of claims 1 to 16, further comprising a first medium supply unit connected to the first flow path. The cell treatment apparatus according to claim 4 or 5, further comprising a second medium supply unit connected to the third flow path. The cell treatment apparatus according to any one of claims 1 to 18, further comprising a cleaning liquid supply unit that supplies a cleaning liquid to the cell supply unit.

Images