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WO1999007742A1 - Process and device for producing growth factors - Google Patents

Process and device for producing growth factors Download PDF

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Publication number
WO1999007742A1
WO1999007742A1 PCT/EP1998/004520 EP9804520W WO9907742A1 WO 1999007742 A1 WO1999007742 A1 WO 1999007742A1 EP 9804520 W EP9804520 W EP 9804520W WO 9907742 A1 WO9907742 A1 WO 9907742A1
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WIPO (PCT)
Prior art keywords
storage container
growth factors
platelet concentrate
blood
component separation
Prior art date
Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
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PCT/EP1998/004520
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German (de)
French (fr)
Inventor
Ingo Flesch
Volker Brand
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Individual
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Individual
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Filing date
Publication date
Application filed by Individual filed Critical Individual
Priority to AU89779/98A priority Critical patent/AU8977998A/en
Priority to EP98941379A priority patent/EP1001988A1/en
Priority to US09/463,476 priority patent/US6325829B1/en
Publication of WO1999007742A1 publication Critical patent/WO1999007742A1/en
Anticipated expiration legal-status Critical
Ceased legal-status Critical Current

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    • CCHEMISTRY; METALLURGY
    • C07ORGANIC CHEMISTRY
    • C07KPEPTIDES
    • C07K14/00Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
    • C07K14/435Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from animals; from humans
    • C07K14/475Growth factors; Growth regulators
    • C07K14/49Platelet-derived growth factor [PDGF]

Definitions

  • the invention relates to a method for producing autologous, thrombocytic growth factors (growth factors). Furthermore, the invention relates to a device for producing autologous, thrombocytic growth factors (growth factors) and a system as part of a device for producing autologous thrombocytic growth factors (growth factors).
  • Wound healing of tissue defects on a body outer surface is usually controlled by the body in a natural way with the help of growth factors.
  • the body's own wound healing processes are often not sufficient the body's wound healing, for example, be chronically disturbed as a result of diabetes, so that wounds cannot heal naturally
  • the present invention is based on the problem of providing a method and a device for producing autologous thrombocytic growth factors (growth factors), wound growth being able to be stimulated with the aid of the produced growth factors. Furthermore, a system is intended as a component of a device for producing autologous thrombocytic Growth factors are provided to solve this problem, the method according to the invention is characterized by the following measures
  • a platelet concentrate is obtained from blood
  • the platelet concentrate is then degranulated, in particular by introducing thrombin and calcium into the platelet concentrate, a platelet agglomerate and a solution being formed,
  • the entire method is preferably carried out immediately after the blood is drawn as a bed-si de method. This also ensures that high concentrations of autologous thrombocytic growth factors can be produced or obtained
  • the device according to the invention for producing autologous, thrombocytic growth factors (growth factors) is characterized by
  • the device according to the invention enables the production or extraction of high concentrations of autologous, thrombocytic growth factors (growth factors).
  • the system according to the invention as a component of a device for producing autologous, thrombocytic growth factors is characterized by at least two storage containers, with a first storage container being assigned at least one inlet member and a second storage container being assigned at least one outlet member, the two storage containers being connected to one another by connecting lines and the system being connectable to a blood component separation system via a connecting member
  • the system according to the invention is of a particularly simple construction. It can be produced particularly cost-effectively. Furthermore, the system according to the invention can be used in connection with known and widely used blood component separation systems. Hospitals or the like are accordingly able at low cost to independently autologous, thrombocytic growth factors (growth factors ) to produce
  • FIG. 1 shows a system as part of a device for producing autologous, thrombocytic growth factors
  • the system shown in the drawing is part of a device for producing autologous, thrombocytic growth factors or autologous growth factors from whole blood With the help of the manufactured or obtained autologous groth factors, wound healing processes can be positively influenced.
  • FIG. 1 A system 10 as part of a device for producing autologous growth factors is shown in FIG. 1. With the aid of a connecting element 11, the system 10 can be connected to a blood component separation system, not shown.
  • This blood component separation system can be, for example, that of the company DIDECO S.p.A. Manufacture the autotransfusion device of the Compact-A series. However, any other blood component separation system can also be used
  • the system 10 has two storage containers 12, 13.
  • the storage containers 12, 13 are designed as bags.
  • the storage containers 12, 13 are connected on the one hand to one another and on the other hand to the blood component separation system (not shown).
  • the system 10 has connecting lines 14, 15.
  • the connecting lines 14, 15 are tubes.
  • one of the connecting lines 14 and 15 is connected via a first end 16 and 17 to one of the storage containers 12 and 13, respectively.
  • the connecting lines 14, 15 act on connection points 18, 19 of the storage containers 12, 13.
  • the connecting lines 14 and 15 are attached to a common connecting member 22, the connecting member 11 also engaging at one end 23 on the connecting member 22.
  • the connecting member 22 of the system 10 can be connected to the blood component separation system. The end 24 is closed when the system 10 is not in use 1 by a cap 25
  • the first storage container 12 of the system 10 has an inlet member 26. With the aid of the inlet member 26, substances can be introduced into the storage container 12 in a targeted and contamination-free manner.
  • the inlet member 26 comprises a connection member 27 as a connection to the storage container 12 and a 3-way arranged on the connection member 27. Tap 28 On the 3-way tap 28 is one Syringe 29 can be used, which contains the substances to be introduced into the storage container 12
  • the second reservoir 13 of the system 10 has an outlet member 30. With the aid of the outlet member 30, substances can be removed from the reservoir 13 in a targeted and contamination-free manner.
  • the outlet member 30, like the inlet member 26, has a connector 31 as a connection to the reservoir 13 and one on the connector 31 arranged 3-way tap 32 A syringe or the like, not shown, can be attached to the 3-way tap 32, by means of which the substances can be removed from the storage container 13
  • Both the storage container 12 and the storage container 13 each have a further connecting element 33, 34.
  • the connecting element 33, 34 are so-called retrans-adapters
  • a further connecting line 35 engages on the connecting member 22 in addition to the connecting lines 14, 15 and the connecting member 11.
  • An additional not shown inlet member can be attached to the connecting line 35 in order to introduce further south punches into the system 10 in a contamination-free manner Connection line 35 closed by a cap 36
  • the connecting lines ⁇ L 15, 35 are each assigned a clamping member 37, 38, 39, with the help of the clamping members 37, 38, 39 the flow direction within the system 10 can be determined
  • the blood namely whole blood immediately after blood collection or wa h rend of the non dargestel] th Blood component separation system supplied
  • the blood component separation system works on the principle of a centrifuge.
  • the blood component separation system extracts a platelet concentrate from the blood.
  • the platelet concentrate is supplied from the blood component separation system shown to the first reservoir 12 via the connecting line 14.
  • the clamping member 37 is opened
  • the clamping organs 38, 39 are closed. Unnecessary blood components such as erythrocytes, leukocytes and plasma are retransferred by the blood component separation system
  • the relevant specialist is familiar with the mode of operation and parameterization of the blood component separation system for obtaining the platelet concentrate.
  • whole blood at 350 * g g corresponds to gravitational acceleration
  • g corresponds to gravitational acceleration
  • the platelet-rich plasma can then be centrifuged at 1370 * g for 15 minutes. Platelets are pelleted. Surplus plasma is again discarded.
  • the desired platelet concentrate remains
  • the thrombocyte concentrate located in the first storage container 12 is then added, via the inlet member 26, a thrombi-calcium mixture located in the syringe 29.
  • the 3-way valve 28 is opened, the clamping member 37 is closed and the thrombin-calcium mixture is mixed is injected This takes place at room temperature
  • the thrombin-calcium mixture is a standard preparation according to the German Pharmacy Book (DAB).
  • DAB German Pharmacy Book
  • a dose of 5000 IU thrombin and 10 ml calcium 10% per 500 ml whole blood is used.
  • a variation of the thrombin concentration is possible, to achieve a different consistency (liquid to gel-like) of the growth factor mixture
  • the thrombin-calcium mixture accordingly has the following composition
  • thrombi-calcium mixture it is also conceivable to vary the thrombi-calcium mixture or to use other mixtures to degranulate the platelet concentrate. 1000 IU thrombin and 10 ml calcium 10% per 500 ml whole blood can also be used
  • the platelets in the platelet concentrate degranulate and release the so-called granules with the autologous growth factors located therein.
  • the platelets agglomerate to form a platelet agglomerate in the released autologous growth factors it is about
  • Fibroblast Growth Factors Pl at 1 et De ⁇ ved Growth Factors, Transforming Growth Factors, Fibronectin
  • the mixture of platelet concentrate and thrombi n-cal ci um mixture is transferred from the first storage container 12 to the second storage container 13.
  • the clamping member 37 and the clamping member 38 are opened.
  • the platelet aggregate is separated from the solution containing the autologous growth factors
  • the platelet agglomerate is discarded
  • the solution located in the storage container 13 is removed from the storage container 13 via the outlet member 30 to form portions.
  • the 3-way valve 1 32 is opened, to which a portion container or the like was previously attached. This is carried out until the entire in Storage container 13 located solution is divided into portions
  • the portioned solution is snap-frozen to a temperature of -40 ° C
  • the solution obtained in this way is mixed with homologous groth factors and / or buffer solution and / or dilution solution, and only afterwards this mixture is portioned and shock-frozen in order to admix the homologous groth factors and / or buffer solution and / or dilution solution is an inlet element, not shown, which can be connected to the connecting line 35
  • the shock-frozen portions can be thawed by the patient.
  • the portions can be applied to the wound daily to stimulate the wound healing process
  • the method according to the invention can also be carried out in a device with a storage container.
  • the platelet concentrate is mixed in one storage container with the thrombi-calzi um mixture and after agglomeration and granulation, the platelet agglomerate is simply clamped off separated from the solution containing the autologous growth factors in this one storage container

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  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Organic Chemistry (AREA)
  • Biochemistry (AREA)
  • Gastroenterology & Hepatology (AREA)
  • Zoology (AREA)
  • Biophysics (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Medicinal Chemistry (AREA)
  • Molecular Biology (AREA)
  • Proteomics, Peptides & Aminoacids (AREA)
  • Toxicology (AREA)
  • Medicines Containing Material From Animals Or Micro-Organisms (AREA)

Abstract

A process and device are disclosed for producing autologous, thrombocytic growth factors, as well as a system which is part of the device for producing autologous growth factors. According to the disclosed device for producing autologous growth factors, a thrombocyte concentrate is extracted from blood and a defined amount of thrombin and calcium is then added to the thrombocyte concentrate. The resultant thrombocyte agglomerate is separated from a solution which contains the growth factors, and the thus obtained solution is portioned and frozen.

Description

Verfahren und Vorrichtung zur Herstellung von Wachstumsfaktoren. Method and device for producing growth factors.

Die Erfindung betrifft ein Verfahren zur Herstellung autologer, thrombozytarer Wachstumsfaktoren (Growth Factors) Des weiteren betrifft die Erfindung eine Vorrichtung zur Herstellung autologer, thrombozytarer Wachstumsfaktoren (Growth Factors) sowie ein System als Bestandteil einer Vorrichtung zur Herstellung autologer thrombozytarer Wachstumsfaktoren (Growth Factors)The invention relates to a method for producing autologous, thrombocytic growth factors (growth factors). Furthermore, the invention relates to a device for producing autologous, thrombocytic growth factors (growth factors) and a system as part of a device for producing autologous thrombocytic growth factors (growth factors).

Die Wundheilung von Gewebsdefekten an einer Korperaußenf1 ache , insbesondere von Schürfwunden, Rißwunden oder dergleichen, wird üblicherweise vom Korper auf natürliche Weise mit Hilfe von Wachstumsfaktoren (Growth Factors) gesteuert Bei besonders großflächigen Gewebsdefekten hingegen sind häufig die körpereigenen Wundhei 1 ungsprozesse nicht ausreichend Auch kann die körpereigene Wundheilung z B in Folge von Zuckerkrankheit chronisch gestört sein, so daß auf natürliche Weise Wunden nicht heilen könnenWound healing of tissue defects on a body outer surface, in particular abrasions, lacerations or the like, is usually controlled by the body in a natural way with the help of growth factors. In the case of particularly large tissue defects, however, the body's own wound healing processes are often not sufficient the body's wound healing, for example, be chronically disturbed as a result of diabetes, so that wounds cannot heal naturally

Hiervon ausgehend liegt der vorliegenden Erfindung das Problem zugrunde, ein Verfahren sowie eine Vorrichtung zur Herstellung autologer thrombozytarer Wachstumsfaktoren (Growth Factors) bereitzustellen, wobei mit Hilfe der hergestellten Growth Factors die Wundheilung stimuliert werden kann Ferner soll ein System als Bestandteil einer Vorrichtung zur Herstellung autologer thrombozytarer Wachstumsfaktoren (Growth Factors) bereitgestellt werden Zur Losung dieses Problems ist das erfindungsgemaße Verfahren durch folgende Maßnahmen gekennzeichnetProceeding from this, the present invention is based on the problem of providing a method and a device for producing autologous thrombocytic growth factors (growth factors), wound growth being able to be stimulated with the aid of the produced growth factors. Furthermore, a system is intended as a component of a device for producing autologous thrombocytic Growth factors are provided To solve this problem, the method according to the invention is characterized by the following measures

a) aus Blut wird ein Thrombozytenkonzentrat gewonnen,a) a platelet concentrate is obtained from blood,

b) das Thrombozytenkonzentrat wird sodann degranul lert , insbesondere durch Einbringen von Thrombin und Calzium in das Thrombozytenkonzentrat, wobei ein Thrombozyten- agglomerat und eine Losung entsteht,b) the platelet concentrate is then degranulated, in particular by introducing thrombin and calcium into the platelet concentrate, a platelet agglomerate and a solution being formed,

c) das Thrombozytenagglomerat wird von der Losung getrennt,c) the platelet agglomerate is separated from the solution,

d) darauffolgend wird die Losung portioniert und gefrorend) the solution is then portioned and frozen

Mit dem erfi ndungsgemaßen Verfahren sind auf einfache und kostengünstige Weise hohe Konzentrationen an autologen, throm- bozytaren Wachstumsfaktoren (Growth Factors) herstellbar bzw gewinnbar Durch die Verwendung derselben laßt sich auch bei großflächigen Gewebsdefekten oder bei chronisch gestörter Wund- heilung ein guter Wundhei lungsprozeß erzielenWith the method according to the invention, high concentrations of autologous, thrombocytic growth factors can be produced or obtained in a simple and cost-effective manner. By using the same, a good wound healing process can also be achieved in the case of large-area tissue defects or chronically disturbed wound healing

Vorzugsweise wird das gesamte Verfahren im unmittelbaren Anschluß an die Blutentnahme als bed-si de-Verfahren durchgeführt Auch dies gewährleistet, daß hohe Konzentrationen an autologen thrombozytaren Wachstumsfaktoren (Growth Factors) herstellbar bzw gewinnbar sindThe entire method is preferably carried out immediately after the blood is drawn as a bed-si de method. This also ensures that high concentrations of autologous thrombocytic growth factors can be produced or obtained

Die erfindungsgemaße Vorrichtung zur Herstellung autologer, thrombozytarer Wachstumsfaktoren (Growth Factors) ist gekenn- zeichnet durchThe device according to the invention for producing autologous, thrombocytic growth factors (growth factors) is characterized by

a) ein Bl utkomponenten-Separationssytem,a) a blood component separation system,

b) mindestens einen mit dem Bl utkomponenten-Separationssytem verbundenen Vorratsbehalte ,b) at least one storage container connected to the blood component separation system,

c) mindestens ein dem Vorratsbehalter zugeordnetes Einlaßorgan, d) mindestens ein Auslaßorganc) at least one inlet member assigned to the storage container, d) at least one outlet member

Die erfindungsgemaße Vorrichtung ermöglicht die Herstellung bzw die Gewinnung hoher Konzentrationen an autologen, thrombo- zytaren Wachstumsfaktoren (Growth Factors)The device according to the invention enables the production or extraction of high concentrations of autologous, thrombocytic growth factors (growth factors).

Das erfindungsgemaße System als Bestandteil einer Vorrichtung zur Herstellung autologer, thrombozytarer Wachstumsfaktoren (Growth Factors) ist gekennzeichnet durch mindestens zwei Vorratsbehalter, wobei einem ersten Vorratsbehalter mindestens ein Einlaßorgan und einem zweiten Vorratsbehalter mindestens ein Auslaßorgan zugeordnet ist, wobei die beiden Vorratsbehalter durch Verbindungsleitungen miteinander verbunden sind und wobei das System über ein Anschlußorgan an ein Blutkomponenten- Separationssystem anschließbar istThe system according to the invention as a component of a device for producing autologous, thrombocytic growth factors is characterized by at least two storage containers, with a first storage container being assigned at least one inlet member and a second storage container being assigned at least one outlet member, the two storage containers being connected to one another by connecting lines and the system being connectable to a blood component separation system via a connecting member

Das erfindungsgemaße System ist von besonders einfachem Aufbau Es ist besonders kostengustig herstellbar Ferner ist das erfindungsgemaße System im Zusammenhang mit bekannten und vielfaltig verbreiteten Blutkomponenten-Separationssystemen einsetzbar Krankenhauser oder dergleichen werden demnach mit geringem Kostenaufwand in die Lage versetzt, selbsstandig autologe, thrombozytare Wachstumsfaktoren (Growth Factors) herzustel lenThe system according to the invention is of a particularly simple construction. It can be produced particularly cost-effectively. Furthermore, the system according to the invention can be used in connection with known and widely used blood component separation systems. Hospitals or the like are accordingly able at low cost to independently autologous, thrombocytic growth factors (growth factors ) to produce

Bevorzugte Weiterbildungen der Erfindung ergeben sich aus den Unteranspruchen und der Beschreibung Anhand der Zeichnung wird ein Ausfuhrungsbeispiel der Erfindung naher erläutert In der Zeichnung zeigtPreferred developments of the invention result from the subclaims and the description. An exemplary embodiment of the invention is explained in more detail with the aid of the drawing

Fig 1 Ein System als Bestandteil einer Vorrichtung zur Herstellung autologer, thrombozytarer Wachstumsfaktoren (Growth Factors)1 shows a system as part of a device for producing autologous, thrombocytic growth factors

Das in der Zeichnung dargestellte System ist ein Bestandteil einer Vorrichtung zur Herstellung autologer, thrombozytarer Wachstumsfaktoren bzw autologer Growth Factors aus Vollblut Mit Hilfe der hergestellten bzw. gewonnenen autologen Groth Factors sind Wundhei lungsprozeße positiv beeinflußbar.The system shown in the drawing is part of a device for producing autologous, thrombocytic growth factors or autologous growth factors from whole blood With the help of the manufactured or obtained autologous groth factors, wound healing processes can be positively influenced.

Ein System 10 als Bestandteil einer Vorrichtung zur Herstellung autologer Growth Factors zeigt Fig. 1. Mit Hilfe eines Anschlußorgans 11 ist das System 10 an ein nicht dargestelltes Blutkomponenten-Separationssystem anschließbar. Bei diesem Blutkomponenten-Separationssystem kann es sich zum Beispiel um das von der Firma DIDECO S.p.A. hergestellte Autotransfusionsgerät der Baureihe Compact-A handeln Es sind aber auch beliebig andere Blutkomponenten-Separationssysteme einsetzbarA system 10 as part of a device for producing autologous growth factors is shown in FIG. 1. With the aid of a connecting element 11, the system 10 can be connected to a blood component separation system, not shown. This blood component separation system can be, for example, that of the company DIDECO S.p.A. Manufacture the autotransfusion device of the Compact-A series. However, any other blood component separation system can also be used

Das System 10 verfugt über zwei Vorratsbehalter 12, 13. Die Vorratsbehalter 12, 13 sind als Beutel ausgebildet. Die Vor- ratsbehalter 12, 13 sind einerseits untereinander und andererseits mit dem nicht dargestellten Blutkomponenten-Separationssystem verbunden Hierzu verfugt das System 10 über Verbindungsleitungen 14, 15. Bei den Verbindungsleitungen 14, 15 handelt es sich um Schlauche.The system 10 has two storage containers 12, 13. The storage containers 12, 13 are designed as bags. The storage containers 12, 13 are connected on the one hand to one another and on the other hand to the blood component separation system (not shown). For this purpose, the system 10 has connecting lines 14, 15. The connecting lines 14, 15 are tubes.

Gemäß Fig. 1 ist jeweils eine der Verbindungsleitungen 14 bzw. 15 über ein erstes Ende 16 bzw. 17 mit einem der Vorratsbehalter 12 bzw 13 verbunden. Die Verbindungsleitungen 14, 15 greifen hierzu an Anschl ußorgenen 18, 19 der Vorratsbehalter 12, 13 an. Mit zweiten Enden 20 bzw. 21 sind die Verbindungsleitungen 14 bzw. 15 an einem gemeinsamen Verbindungsorgan 22 angebracht, wobei an dem Verbindungsorgan 22 ebenfalls das Anschlußorgan 11 mit einem Ende 23 angreift. Mit dem zum Ende 23 gegenüberliegenden Ende 24 hingegen ist das Verbindungsorgan 22 des Systems 10 am Blutkomponenten-Separationssystem anschließbar. Das Ende 24 ist im Nichtgebrauchsfal 1 des Systems 10 durch eine Kappe 25 verschlossen1, one of the connecting lines 14 and 15 is connected via a first end 16 and 17 to one of the storage containers 12 and 13, respectively. For this purpose, the connecting lines 14, 15 act on connection points 18, 19 of the storage containers 12, 13. With second ends 20 and 21, the connecting lines 14 and 15 are attached to a common connecting member 22, the connecting member 11 also engaging at one end 23 on the connecting member 22. By contrast, with the end 24 opposite the end 23, the connecting member 22 of the system 10 can be connected to the blood component separation system. The end 24 is closed when the system 10 is not in use 1 by a cap 25

Der erste Vorratsbehalter 12 des Systems 10 verfugt über ein Einlaßorgan 26. Mit Hilfe des Einlaßorgans 26 sind in den Vorratsbehalter 12 gezielt und kontaminationsfrei Substanzen einbringbar Das Einlaßorgan 26 umfaßt ein Anschlußorgan 27 als Verbindung zum Vorratsbehalter 12 sowie einen am Anschlußorgan 27 angeordneten 3-Wege-Hahn 28 Am 3-Wege-Hahn 28 ist eine Spritze 29 ansetzbar, die die in den Vorratsbehalter 12 einzubringenden Substanzen enthaltThe first storage container 12 of the system 10 has an inlet member 26. With the aid of the inlet member 26, substances can be introduced into the storage container 12 in a targeted and contamination-free manner. The inlet member 26 comprises a connection member 27 as a connection to the storage container 12 and a 3-way arranged on the connection member 27. Tap 28 On the 3-way tap 28 is one Syringe 29 can be used, which contains the substances to be introduced into the storage container 12

Der zweite Vorratsbehalter 13 des Systems 10 verfugt über ein Auslaßorgan 30 Mit Hilfe des Auslaßorgans 30 sind aus dem Vorratsbehalter 13 gezielt und kontaminationsfrei Substanzen entnehmbar Das Auslaßorgan 30 verfugt ebenso wie das Einlaßorgan 26 über ein Anschlußorgan 31 als Verbindung zum Vorratsbehalter 13 sowie über einen am Anschlußorgan 31 angeordneten 3-Wege- Hahn 32 Am 3-Wege-Hahn 32 ist eine nicht dargestellte Spritze oder dergleichen ansetzbar, mit Hilfe derer die Substanzen aus dem Vorratsbehalter 13 entnehmbar sindThe second reservoir 13 of the system 10 has an outlet member 30. With the aid of the outlet member 30, substances can be removed from the reservoir 13 in a targeted and contamination-free manner. The outlet member 30, like the inlet member 26, has a connector 31 as a connection to the reservoir 13 and one on the connector 31 arranged 3-way tap 32 A syringe or the like, not shown, can be attached to the 3-way tap 32, by means of which the substances can be removed from the storage container 13

Sowohl der Vorratsbehalter 12 als auch der Vorratsbehalter 13 verfugen über jeweils ein weiteres Anschlußorgan 33, 34 Bei den Ansch ' ußorgan 33, 34 handelt es sich um sogenannte Retrans- AdapterBoth the storage container 12 and the storage container 13 each have a further connecting element 33, 34. The connecting element 33, 34 are so-called retrans-adapters

Gemäß Fig 1 greift am Verbindungsorgan 22 neben den Verbindungsleitungen 14, 15 und dem Anschlußorgan 11 eine weitere Verbi ndungsl eitung 35 An der Verbi ndungsleitung 35 kann ein weiters nicht dargesteltes Einlaßorgan angebracht werden um weitere SuDstanzen in das System 10 kontaminationsfrei einzubringen _m Nichtgebrauchsfal 1 ist die Verbindungsl eitung 35 durch ein Kappe 36 verschlossenAccording to FIG. 1, a further connecting line 35 engages on the connecting member 22 in addition to the connecting lines 14, 15 and the connecting member 11. An additional not shown inlet member can be attached to the connecting line 35 in order to introduce further south punches into the system 10 in a contamination-free manner Connection line 35 closed by a cap 36

Den Verbindungsleitungen \L 15, 35 ist jeweils ein Klemmorgan 37, 38, 39 zugeordnet wobei mit Hilfe der Klemmorgane 37, 38, 39 die Flußπchtung innerhalb des Systems 10 bestimmt werden kannThe connecting lines \ L 15, 35 are each assigned a clamping member 37, 38, 39, with the help of the clamping members 37, 38, 39 the flow direction within the system 10 can be determined

Die Arbeitsweise der erfindungsgemaßen Vorrichtung zur Herstellung autologer Growth Factors sowie des erfindungsgemaßen Systems IC als Bestandteil der erfindungsgemaßen Vorrichtung und damit aas erfindungsgemaße Verfahren werden nachfolgend erl autertThe mode of operation of the device according to the invention for producing autologous growth factors and of the system IC according to the invention as a component of the device according to the invention and thus of the method according to the invention are explained below

Das Blut nämlich Vollblut wird unmittelbar nach beziehungsweise wahrend der Blutentnahme dem nicht dargestel ] ten Blutkomponenten-Separationssyte zugeführt Das Blutkompo- nenten-Separationssystem arbeitet nach dem Prinzip einer Zentrifuge Das Bl utkomponenten-Separationssytem gewinnt aus dem Blut ein Thrombozytenkonzentrat Das Thrombozytenkonzentrat wird vom dargestellten Blutkomponenten-Separationssystem über die Verbindungsleitung 14 dem ersten Vorratsbehalter 12 zugeführt Hierzu ist das Klemmorgan 37 geöffnet, die Klemmorgane 38, 39 hingegen sind geschlossen Nicht benotigte Bl utbestandtei 1 e wie zum Beispiel Erythrozyten , Leukozyten, Plasma werden vom Blutkomponenten-Separationssystem rucktrans- fundiertThe blood namely whole blood immediately after blood collection or wa h rend of the non dargestel] th Blood component separation system supplied The blood component separation system works on the principle of a centrifuge. The blood component separation system extracts a platelet concentrate from the blood. The platelet concentrate is supplied from the blood component separation system shown to the first reservoir 12 via the connecting line 14. For this purpose, the clamping member 37 is opened The clamping organs 38, 39, on the other hand, are closed. Unnecessary blood components such as erythrocytes, leukocytes and plasma are retransferred by the blood component separation system

Arbeitsweise und Parametπerung des Bl utkomponenten-Separa- tionssystems zur Gewinnung des Thrombozytenkonzentrats sind dem einschlagigen Fachmann geläufig So kann zum Beispiel das Vollblut bei 350 * g (g entspricht der Erdbeschleunigung) für 6 Minuten zentπ fugiert werden Hierbei entsteht ein pl attchenreiches Plasma als Überstand, die entstehende Erythrozytenfraktion wird verworfen Das pl attchenreiche Plasma kann sodann bei 1370 * g für 15 Minuten zentπ fugiert werden Dabei werden Thrombozyten pelletiert Überstehendes Plasma wird wiederum verworfen Das gewünschte Thrombozytenkonzentrats bleibt dabei übrigThe relevant specialist is familiar with the mode of operation and parameterization of the blood component separation system for obtaining the platelet concentrate. For example, whole blood at 350 * g (g corresponds to gravitational acceleration) can be centrifuged for 6 minutes. This produces a platelet-rich plasma as a supernatant, the resulting erythrocyte fraction is discarded. The platelet-rich plasma can then be centrifuged at 1370 * g for 15 minutes. Platelets are pelleted. Surplus plasma is again discarded. The desired platelet concentrate remains

Dem im ersten Vorratsbehalter 12 befindlichen Thrombozytenkonzentrat wird anschließend über das Einlaßorgan 26 eine in der Spritze 29 befindliche Thrombi n-Calzium-Mischung zugesetzt Hierzu wird das 3-Wege-Ventι 1 28 geöffnet das Klemmorgan 37 geschlossen und die Thrombin-Calzi u -Mi schung wird einge- spritzt Dies erfolgt bei RaumtemperaturThe thrombocyte concentrate located in the first storage container 12 is then added, via the inlet member 26, a thrombi-calcium mixture located in the syringe 29. For this purpose, the 3-way valve 28 is opened, the clamping member 37 is closed and the thrombin-calcium mixture is mixed is injected This takes place at room temperature

Bei der Thrombin-Cal zium-Mischung handelt es sich um eine Stan- dardpraparation gemäß dem Deutschen Apothekenbuch (DAB) Es wird eine Dosis von 5000 IE Thrombin und 10 ml Calzium 10% pro 500ml Vollblut verwendet Eine Variation der Thrombinkonzen- tration ist möglich, um eine unterschiedliche Konsistenz (flussig bis gelartig) des Wachstumsfaktoren-Gemisches zu erzielen Die Thrombin-Calzium-Mischung weist demnach folgende Zusammensetzung aufThe thrombin-calcium mixture is a standard preparation according to the German Pharmacy Book (DAB). A dose of 5000 IU thrombin and 10 ml calcium 10% per 500 ml whole blood is used. A variation of the thrombin concentration is possible, to achieve a different consistency (liquid to gel-like) of the growth factor mixture The thrombin-calcium mixture accordingly has the following composition

5000 IE Thrombin,5000 IU thrombin,

10 ml Calzium-Losung 10%,10 ml calcium solution 10%,

Aus 500 ml Blut (Vollblut) können in etwa 20ml bis 40 ml Thrombozytenkonzentrat gewonnen werden, die im Vorratsbehalter 12 mit der obigen Thrombin-Calci um-Mi schung vermischt werdenFrom 500 ml of blood (whole blood), about 20 ml to 40 ml of platelet concentrate can be obtained, which are mixed in the storage container 12 with the above thrombin-calcium mixture

Es ist auch denkbar die Thrombi n-Cal ziu -Mi schung zu variieren bzw andere Mischungen zur Degranul lerung des Thrombozytenkonzentrat einzusetzen So können auch 1000 IE Thrombin und 10 ml Calzium 10% pro 500ml Vollblut verwendet werdenIt is also conceivable to vary the thrombi-calcium mixture or to use other mixtures to degranulate the platelet concentrate. 1000 IU thrombin and 10 ml calcium 10% per 500 ml whole blood can also be used

Durch das Einbringen der Thro bin-Cal ci u -Mischung in das Thrombozytenkonzentrat degranul leren die Thrombozyten des Thrombozytenkonzentrats und setzen die sogenannte Granula mit den darin lokalisierten autologen Growth Factors frei Darüber hinaus agglomerieren die Thrombozyten zu einem Thrombozyten- agglomerat Bei den freigesetzten autologen Growth Factors handelt es sich umBy introducing the Thro bin-Cal ci u mixture into the platelet concentrate, the platelets in the platelet concentrate degranulate and release the so-called granules with the autologous growth factors located therein. In addition, the platelets agglomerate to form a platelet agglomerate in the released autologous growth factors it is about

Fibroblast Growth Factors, Pl at 1 et Deπved Growth Factors, Transforming Growth Factors, FibronectinFibroblast Growth Factors, Pl at 1 et Deπved Growth Factors, Transforming Growth Factors, Fibronectin

Die Mischung aus Thrombozytenkonzentrat und Thrombi n-Cal ci um- Mischung wird vom ersten Vorratsbehalter 12 in den zweiten Vorratsbehalter 13 überfuhrt Hierzu wird das Klemmorgan 37 sowie das Klemmorgan 38 geöffnet Hierbei wird das Thrombozyten- agglo erat von der die autologen Growth Factors enthaltenden Losung getrennt Das Thrombozytenaggl omerat wird verworfen Die im Vorratsbehalter 13 befindliche Losung wird unter Bildung von Portionen aus dem Vorratsbehalter 13 über das Auslaßorgan 30 entnommen Hierzu wird das 3-Wege-Ventι 1 32 geöffnet, an dem zuvor ein Portiongefaß oder dergleichen angebracht wurde Dies wird solange durchgeführt, bis die gesamte im Vorratsbehalter 13 befindliche Losung in Portionen aufgeteilt istThe mixture of platelet concentrate and thrombi n-cal ci um mixture is transferred from the first storage container 12 to the second storage container 13. For this purpose, the clamping member 37 and the clamping member 38 are opened. Here, the platelet aggregate is separated from the solution containing the autologous growth factors The platelet agglomerate is discarded The solution located in the storage container 13 is removed from the storage container 13 via the outlet member 30 to form portions. For this purpose, the 3-way valve 1 32 is opened, to which a portion container or the like was previously attached. This is carried out until the entire in Storage container 13 located solution is divided into portions

Unmittelbar im Anschluß an das Portionieren wird die portionierte Losung auf eine Temperatur von -40°C schockgefrorenImmediately after portioning, the portioned solution is snap-frozen to a temperature of -40 ° C

Das gesamte Verfahren wird als bed-side-Verfahren durchgeführtThe entire process is carried out as a bed-side process

Auch ist es möglich, daß im Anschluß an das Einbringen der Thrombin-Calzium-Mischung in das Thrombozytenkonzentrat und das Abtrennen des Thro bozytenagglomerats die hierbei gewonnene Losung mit homologen Groth Factors und/oder Pufferlosung und/oder Verdunnungslosung vermischt wird, und daß erst im Anschluß hieran diese Mischung portioniert und schockgefroren wird Zum Zumischen der homologen Groth Factors und/oder Pufferlosung und/oder Verdunnungslosung dient ein nicht dargesteltes Einlaßorgan, das an der Verbindungsleitung 35 angeschlossen werden kannIt is also possible that, after the thrombin-calcium mixture has been introduced into the platelet concentrate and the thrombocyte agglomerate has been separated off, the solution obtained in this way is mixed with homologous groth factors and / or buffer solution and / or dilution solution, and only afterwards this mixture is portioned and shock-frozen in order to admix the homologous groth factors and / or buffer solution and / or dilution solution is an inlet element, not shown, which can be connected to the connecting line 35

Die schockgefrorenen Portionen können vom Patienten aufgetaut werden Zur Stimulierung des Wundhei 1 ungsprozesses können die Portionen täglich auf die Wunde aufgetragenThe shock-frozen portions can be thawed by the patient. The portions can be applied to the wound daily to stimulate the wound healing process

Auch soll darauf hingewiesen werden, daß das erfindungsgemaße Verfahren ebenso in einer Vorrichtung mit einem Vorratsbehalter durchgeführt werden kann Hierbei wird das Thrombozytenkonzentrat in dem einen Vorratsbehalter mit der Thrombi n-Calzi um- Mischung gemischt und nach dem Agglomerieren und Granulieren wird das Thrombozytenagglomerat durch einfaches Abklemmen von der die autologen Growth Factors enthaltenden Losung in diesem einen Vorratsbehalter getrenntIt should also be pointed out that the method according to the invention can also be carried out in a device with a storage container. Here, the platelet concentrate is mixed in one storage container with the thrombi-calzi um mixture and after agglomeration and granulation, the platelet agglomerate is simply clamped off separated from the solution containing the autologous growth factors in this one storage container

***** Bez uαs zei chen l i te***** Please refer to the left

System 31 AnschlußorganSystem 31 connecting element

Anschl ußorgan 32 3-Wege-Venti 1Connection element 32 3-way valve 1

Vorratsbehalter 33 Anschl ußorganStorage container 33 connecting element

Vorratsbehalter 34 AnschlußorganStorage container 34 connecting member

Verbindungsleitung 35 VerbindungsleitungConnection line 35 Connection line

Verbindungsleitung 36 KappeConnection line 36 cap

Ende 37 KlemmorganEnd 37 clamp member

Ende 38 KlemmorganEnd 38 clamp member

Anschl ußorgan 39 KlemmorganConnecting element 39 clamping element

Anschl ußorganConnecting element

EndeThe End

EndeThe End

VerbindungsorganLiaison body

EndeThe End

EndeThe End

Kappecap

Ei nl aßorgenTake care

Anschl ußorganConnecting element

3-Wege-Ventil3-way valve

Spritzesyringe

Auslaßorgan Outlet organ

Claims

Patentansprüche claims 1. Verfahren zur Herstellung autologer, thrombozytarer Wachstumsfaktoren (Growth Factors), gekennzeichnet durch folgende Maßnahmen:1. A process for producing autologous, thrombocytic growth factors, characterized by the following measures: a) aus Blut wird ein Thrombozytenkonzentrat gewonnen,a) a platelet concentrate is obtained from blood, b) das Thrombozytenkonzentrat wird sodann degranul lert , insbesondere durch Einbringen von Thrombin und Calzium in das Thrombozytenkonzentrat, wobei ein Thrombozyten- agglomerat und eine Losung entsteht,b) the platelet concentrate is then degranulated, in particular by introducing thrombin and calcium into the platelet concentrate, a platelet agglomerate and a solution being formed, das Thrombozytenagglomerat wird von der Losung getrennt,the platelet agglomerate is separated from the solution, darauffolgend wird die hierbei gewonnene Losung portioniert und gefroren.the resulting solution is then portioned and frozen. 2. Verfahren nach Anspruch 1, dadurch gekennzeichnet, daß das Blut zur Gewinnung des Thrombozytenkonzentrats unmittelbar anschließend an eine Blutentnahme einem Blutkomponenten- Separationssytem zugeführt wird.2. The method according to claim 1, characterized in that the blood for obtaining the platelet concentrate is fed to a blood component separation system immediately after a blood sample. 3. Verfahren nach Anspruch 1 oder 2, dadurch gekennzeichnet, daß dem Thrombozytenkonzentrat eine Thrombin-Calzium-Mi schung zugesetzt wird.3. The method according to claim 1 or 2, characterized in that a thrombin-calcium mix is added to the platelet concentrate. 4. Verfahren nach Anspruch 3, dadurch gekennzeichnet, daß Thrombozyten des Thrombozytenkonzentrats beim Zusetzen der Thrombin-Calcium-Mischung degranul leren sowie agglomenren und hierbei Granula freisetzen, wobei in der Granula die autologen Growth Factors lokalisiert sind. 4. The method according to claim 3, characterized in that platelets of the platelet concentrate when adding the thrombin-calcium mixture degranul leren and agglomerate and thereby release granules, the autologous growth factors are localized in the granules. 5. Verfahren nach einem oder mehreren der Ansprüche 1 bis 4, dadurch gekennzeichnet, daß die Thrombin-Calzium-Mischung dem Thrombozytenkonzentrat bei Raumtemperatur zugesetzt wird.5. The method according to one or more of claims 1 to 4, characterized in that the thrombin-calcium mixture is added to the platelet concentrate at room temperature. s 6. Verfahren nach einem oder mehreren der Ansprüche 1 bis 5, dadurch gekennzeichnet, daß die die autologen Growth Factors enthaltene Lösung portioniert und anschließend schockgefroren wird.s 6. The method according to one or more of claims 1 to 5, characterized in that the solution containing the autologous growth factors is portioned and then snap-frozen. o 7. Verfahren nach Anspruch 6, dadurch gekennzeichnet, daß die Losung auf eine Temperatur von -40°C schockgefroren wird.7. The method according to claim 6, characterized in that the solution is snap frozen to a temperature of -40 ° C. 8. Verfahren nach einem oder mehreren der Ansprüche 1 bis 7, dadurch gekennzeichnet, daß im Anschluß an das Einbringen der Thrombin-Calzium-Mischung in das Thrombozytenkonzentrat und das Abtrennen des Thrombozytenagglomerats die hierbei gewonnene Losung mit homologen Groth Factors und/oder Pufferlosung und/oder Verdunnungslosung vermischt wird, und daß erst im Anschluß hieran diese Mischung portioniert und schockgefroren wird.8. The method according to one or more of claims 1 to 7, characterized in that following the introduction of the thrombin-calcium mixture into the platelet concentrate and the separation of the platelet agglomerate, the solution obtained with homologous groth factors and / or buffer solution and / or dilution solution is mixed, and only then is this mixture portioned and snap-frozen. 9. Verfahren nach einem oder mehreren der Ansprüche 1 bis 8, dadurch gekennzeichnet, daß das das gesamte Verfahren im unmittelbaren Anschluß an die Blutentnahme als bed-side- Verfahren durchgeführt wird9. The method according to one or more of claims 1 to 8, characterized in that the entire method is carried out immediately after the blood withdrawal as a bed-side method 10. Vorrichtung zur Herstellung autologer, thrombozytarer Wachstumsfaktoren (Growth Factors), gekennzeichnet durch-10. Device for producing autologous, thrombocytic growth factors, characterized by a) ein Bl utkomponenten-Separationssytem,a) a blood component separation system, b) mindestens einen mit αem Bl utkomponenten-Separationssytem verbundenen Vorratsbenalter (12, 13),b) at least one storage age (12, 13) connected to the blood component separation system, c) mindestens ein dem Vorratsbehalter (12) zugeordnetes Einlaßorgan (26),c) at least one inlet member (26) assigned to the storage container (12), d) mindestens ein Ausl aßorgan(30) d) at least one outlet element (30) 11 Vorrichtung nach Anspruch 10, gekennzeichnet durch zwei Vorratsbehalter (12, 13), die einerseits untereinander sowie andererseits mit dem Bl utkomponenten-Separationssytem verbunden sind und ein geschlossenes System bilden11 Device according to claim 10, characterized by two storage containers (12, 13) which are connected on the one hand to one another and on the other hand to the blood component separation system and form a closed system 12 Vorrichtung nach Anspruch 10 oder 11, dadurch gekennzeichnet, daß das Blutkomponenten-Separationssytem der Gewinnung eines Thrombozytenkonzentrats aus Blut dient12 Device according to claim 10 or 11, characterized in that the blood component separation system is used to obtain a platelet concentrate from blood 13 Vorrichtung nach einem oder mehreren der Ansprüche 10 bis13 Device according to one or more of claims 10 to 12, dadurch gekennzeichnet, daß ein erster Vorratsbehalter (12) einerseits mit dem Bl utkomponenten-Separationssytem und andererseits mit einem zweiten Vorratsbehalter (13) verbunden ist, wobei der erste Vorratsbehalter (12) das gewonnene Thrombozytenkonzentrats aufnimmt und das Einlaßorgan (26) für das Einbringen der Thro bin-Calzi um-Mischung aufweist12, characterized in that a first storage container (12) is connected on the one hand to the blood component separation system and on the other hand to a second storage container (13), the first storage container (12) receiving the platelet concentrate obtained and the inlet member (26) for it Introducing the Thro bin-Calzi um mixture 14 Vorrichtung nach einem oder mehreren der Ansprüche 10 bis14 Device according to one or more of claims 10 to 13, dadurch gekennzeichnet, daß der zweite Vorratsbehalter (13) ebenfalls mit dem Bl utkomponenten-Separationssytem verbunden ist, wobei der zweite Vorratsbehalter (13) die Mischung aus Thrombozytenkonzentrat und Thrombin-Cal zi um-Mischung aufnimmt und das Auslaßorgan (30) für das Portionieren dieser Mischung aufwei st13, characterized in that the second storage container (13) is also connected to the blood component separation system, the second storage container (13) receiving the mixture of platelet concentrate and thrombin-calcium mixture and the outlet member (30) for the Serve this mixture in portions 15 Vorrichtung nach einem oder mehreren der Ansprüche 10 bis15 Device according to one or more of claims 10 to 14, gekennzeichnet durch ein weiteres Einlaßorgan, wobei daß weitere Einlaßorgan einem Verbindungsorgan (22) von erstem Vorratsbehalter (12) und zweitem Vorratsbehalter (13) zugeordnet ist14, characterized by a further inlet member, wherein that further inlet member is assigned to a connecting member (22) of the first storage container (12) and the second storage container (13) 16 System als Bestandteil einer Vorrichtung zur Herstellung autologer, thrombozytarer Wachstumsfaktoren (Growth Factors), gekennzeichnet durch mindestens zwei Vorratsbehalter (12, 13), wobei einem ersten Vorratsbehalter (12) mindestens ein Einlaßorgan (26) und einem zweiten Vorratsbehalter (13) mindestens ein Auslaßorgan (30) zugeordnet ist, wobei die beiden Vorratsbehalter (12, 13) durch Verbindungsleitungen (14, 15) miteinander verbunden sind und wobei das System über ein Anschlußorgan (11) an ein Blutkomponenten-Separationssyte anschließbar ist.16 System as part of a device for producing autologous, thrombocytic growth factors, characterized by at least two storage containers (12, 13), a first storage container (12) having at least one inlet member (26) and a second storage container (13) at least one Outlet member (30) is assigned, the two storage containers (12, 13) being connected to one another by connecting lines (14, 15) and the system being connected via a Connection member (11) can be connected to a blood component separation system. 17. System nach Anspruch 16 , gekennzeichnet durch eines oder mehrere der Merkmale der Anprüche 11 bis 15.17. System according to claim 16, characterized by one or more of the features of claims 11 to 15. ***** *****
PCT/EP1998/004520 1997-07-22 1998-07-20 Process and device for producing growth factors Ceased WO1999007742A1 (en)

Priority Applications (3)

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AU89779/98A AU8977998A (en) 1997-08-05 1998-07-20 Process and device for producing growth factors
EP98941379A EP1001988A1 (en) 1997-08-05 1998-07-20 Process and device for producing growth factors
US09/463,476 US6325829B1 (en) 1997-07-22 1998-07-20 Cup for a knee-joint prosthesis

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DE1997133899 DE19733899A1 (en) 1997-08-05 1997-08-05 Method and device for producing autologous, thrombocytic growth factors and system as part of a device for producing autologous, thrombocytic growth factors
DE19733899.2 1997-08-05

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Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2221770A1 (en) * 2002-04-19 2005-01-01 Eduardo Anitua Aldecoa METHOD OF PREPARATION OF A COMPOUND FOR THE REGENERATION OF FABRICS.

Families Citing this family (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
DE19960504A1 (en) * 1999-12-15 2001-08-16 Curasan Ag Regenerating agent
DE19960490A1 (en) * 1999-12-15 2001-07-12 Curasan Ag Regenerating agent

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Publication number Priority date Publication date Assignee Title
WO1993025215A1 (en) * 1992-06-05 1993-12-23 Inoteb Device for producing a supernatant of activated thrombocytes, method for implementing the device and supernatant obtained

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US5585007A (en) * 1994-12-07 1996-12-17 Plasmaseal Corporation Plasma concentrate and tissue sealant methods and apparatuses for making concentrated plasma and/or tissue sealant

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Publication number Priority date Publication date Assignee Title
WO1993025215A1 (en) * 1992-06-05 1993-12-23 Inoteb Device for producing a supernatant of activated thrombocytes, method for implementing the device and supernatant obtained

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
ES2221770A1 (en) * 2002-04-19 2005-01-01 Eduardo Anitua Aldecoa METHOD OF PREPARATION OF A COMPOUND FOR THE REGENERATION OF FABRICS.
ES2221770B2 (en) * 2002-04-19 2006-07-16 Eduardo Anitua Aldecoa METHOD OF PREPARATION OF A COMPOUND FOR THE REGENERATION OF FABRICS.

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