Bag set and method for producing platelet lysate

Abstract

To provide a bag set that can efficiently produce platelet lysate with a simple device configuration, and to provide a method for producing platelet lysate.SOLUTION: A bag set 80 includes: a blood recovery section 51 with a puncture needle; a first whole blood bag 70 for storing whole blood; a second whole blood bag 76 connected downstream of the first whole blood bag 70 via a tube 75b; a leukocyte removal filter 74 connected to the tube 75b; a recovery fluid bag 72 connected to the tube 75b between the leukocyte removal filter 74 and the second whole blood bag 76; a platelet recovery bag 62 connected to the tube 75b between the first whole blood bag 70 and the leukocyte removal filter 74; and a platelet lysate bag 66 connected to the platelet recovery bag 62 via a tube 63.SELECTED DRAWING: Figure 16

Description

[Technical Field]

[0001] The present invention relates to a bag set for extracting growth factors contained in platelets and a method for producing a platelet lysate. [Background technology]

[0002] In recent years, clinical applications of cell therapy have been progressing, in which cells extracted from biological tissues are cultured ex vivo, expanded to the required cell number, and then used for treatment. In particular, mesenchymal stem cells, which can differentiate into various cells, are expected to be applied to regenerative medicine such as the reconstruction of bone, blood vessels, and cardiac muscle. To popularize such cell therapy, a cell culture medium that can efficiently culture the required cells is required.

[0003] Conventionally, animal-derived growth factors such as fetal bovine serum (FBS) have been considered essential for cell culture media for mesenchymal stem cells. However, in recent years, growth factors extracted from platelets contained in human blood have been proposed as an alternative to animal-derived growth factors such as FBS (Non-Patent Document 1). These growth factors are derived from intracellular granules of platelets and extracted to the outside of the cells. Since they are obtained by dissolving platelets, they are also called platelet lysates (PL). It has also been reported that platelet lysates have a higher ability to culture mesenchymal stem cells than FBS.

[0004] One known method for preparing such platelet lysates is to freeze and thaw platelets separated from whole blood (WB) to destroy the cell membrane and extract the growth factors contained in the granules outside the cells. [Prior art documents] [Non-patent literature]

[0005] [Non-Patent Document 1] “Human platelet lysate: Replacing fetal bovine serum as a gold standard for human cell propagation?”, Thierry Burnouf, et.al., Biomaterials,Vol.76, January 2016, Pages 371-387. Summary of the Invention [Problem to be solved by the invention]

[0006] The present invention has been made in relation to the production of the above-mentioned platelet lysate, and aims to provide a bag set and a method for producing platelet lysate that can efficiently produce platelet lysate with a simple device configuration. [Means for solving the problem]

[0007] (1) One aspect of the disclosure below is a bag set used in a method for producing a platelet lysate, the bag set comprising: a step of collecting a buffy coat from whole blood; a step of collecting a platelet concentrate by centrifuging the buffy coat; a step of further concentrating the platelet concentrate by centrifuging the platelet concentrate to prepare a concentrated platelet concentrate; a step of freezing and thawing the concentrated platelet concentrate; and a step of centrifuging the concentrated platelet concentrate that has been subjected to the freezing and thawing steps to collect a supernatant as a platelet lysate, the bag set comprising: a blood collection section equipped with a puncture needle; and a bag set for accommodating whole blood flowing in from the blood collection section. a bag set comprising: a first whole blood bag; a second whole blood bag connected to the downstream side of the first whole blood bag via a first tube; a leukocyte removal filter connected to the first tube between the first whole blood bag and the second whole blood bag; a collection solution bag containing a platelet collection solution, connected to the first tube between the leukocyte removal filter and the second whole blood bag; a platelet collection bag connected to the first tube between the first whole blood bag and the leukocyte removal filter; and a platelet lysate bag connected to the platelet collection bag via a second tube.

[0008] (2) The bag set described in item (1) may further include a first valve unit provided in the first tube between the leukocyte removal filter and the first whole blood bag, and a second valve unit provided in the first tube between the leukocyte removal filter and the second whole blood bag.

[0009] (3) In the bag set according to item (1) or (2), a foreign matter removal filter for removing foreign matter may be provided in the second tube connecting the platelet collection bag and the platelet lysate bag.

[0010] (4) Another aspect of the disclosure below is a blood collection unit equipped with a puncture needle, a first whole blood bag for accommodating whole blood flowing in from the blood collection unit, a second whole blood bag connected to the downstream side of the first whole blood bag via a first tube, a leukocyte removal filter connected to the first tube between the first whole blood bag and the second whole blood bag, a collection solution bag connected to the first tube between the leukocyte removal filter and the second whole blood bag and accommodating a platelet collection solution, a platelet collection bag connected to the first tube between the first whole blood bag and the leukocyte removal filter, and a second whole blood bag connected to the platelet collection bag via a second tube. a platelet lysate bag connected to the leukocyte removal filter, the platelet lysate manufacturing method comprising the steps of: storing whole blood collected from the blood collection section in the first whole blood bag; transferring the whole blood stored in the first whole blood bag to the second whole blood bag through the first tube; collecting platelets adsorbed to the leukocyte removal filter into the platelet collection bag by flowing the platelet collection solution from the collection solution bag toward the platelet collection bag; and detaching the platelet collection bag and the platelet lysate bag from the first tube. [Effects of the Invention]

[0011] The bag set and the method for producing platelet lysate according to the above aspects can efficiently produce platelet lysate with a simple device configuration. [Brief explanation of the drawings]

[0012] [Figure 1] 1 is an explanatory diagram showing the overall configuration of a platelet lysate production system according to a first embodiment. [Figure 2] FIG. 1 is a plan view of a concentrate bag set. [Figure 3] FIG. 1 is a plan view of a collection bag set. [Figure 4] FIG. 1 is a cross-sectional view of a centrifuge. [Figure 5] 1 is a flowchart of a method for producing a platelet lysate according to the first embodiment. [Figure 6] FIG. 6 is an explanatory diagram showing the steps up to pooling of the buffy coat in FIG. 5 in the order of steps. [Figure 7] FIG. 1 is an explanatory diagram showing the steps up to collection of platelet concentrate in order. [Figure 8] FIG. 1 is an explanatory diagram showing the steps up to the transfer of platelet concentrate in order of steps. [Figure 9] FIG. 1 is an explanatory diagram showing the steps for preparing a concentrated platelet concentrate in the order of steps. [Figure 10] FIG. 1 is an explanatory diagram showing the steps up to pooling of concentrated platelet concentrate in the order of steps. [Figure 11] FIG. 1 is an explanatory diagram showing the steps up to collection of platelet lysate in the order of steps. [Figure 12] FIG. 10 is a plan view of a concentration bag set according to a second embodiment. [Figure 13] 10A to 10C are explanatory views showing the steps of concentrating and washing a platelet concentrate according to a second embodiment in the order of steps. [Figure 14] FIG. 10 is a block diagram of a bag set according to a third embodiment. [Figure 15] 10 is a flowchart showing a method for producing a platelet lysate according to a third embodiment. [Figure 16] FIG. 10 is a block diagram of a bag set according to a fourth embodiment. [Figure 17] 10 is a flowchart showing a method for producing a platelet lysate according to a fourth embodiment. [Figure 18]FIG. 18A is a block diagram of a bag set according to a modified example of the fourth embodiment, and FIG. 18B is a block diagram of a bag set for recovering platelets from the leukocyte removal filter of FIG. 18A. [Figure 19] FIG. 10 is a block diagram showing the connection state of the bag set during platelet recovery in the fourth embodiment. [Figure 20] FIG. 10 is a block diagram of a bag set according to a fifth embodiment. [Figure 21] 10 is a flowchart showing a method for producing a platelet lysate according to a fifth embodiment. [Figure 22] FIG. 10 is an explanatory view showing the flow of liquid in a bag set in a method for producing a platelet lysate according to a fifth embodiment. DETAILED DESCRIPTION OF THE INVENTION

[0013] Preferred embodiments of the present invention will be described in detail below with reference to the accompanying drawings. The bags used in the following embodiments are provided with connection ports (transfer ports) for connecting tubing, and at least some of these connection ports may include click tips (easily breakable portions) that can be broken or opened.

[0014] (First embodiment) The method for producing platelet lysate of this embodiment is carried out, for example, at a blood center that processes blood collected from donors to produce various blood products, and is carried out using a production system 10 including a centrifuge 18 and a freezer 20 installed at the blood center.

[0015] As shown in FIG. 1, the platelet lysate production system 10 of this embodiment includes a platelet collection set 12, a concentrate bag set 14, a collection bag set 16, a centrifuge 18, and a freezer 20.

[0016] The platelet collection set 12 is provided as a disposable product and is used to extract platelet concentrate (PC) by removing leukocytes from the buffy coat (BC) obtained by centrifuging whole blood (WB). The platelet collection set 12 includes a BC separation bag 12a, a leukocyte removal filter 12b, and a PC collection bag 12c.

[0017] The BC separation bag 12a is provided with a transfer tube 12d and a transfer tube 12e. As shown in Fig. 6, the transfer tube 12d is a tube connected to a branching portion 12g via a clamp 12f, and is connected to a plurality of BC bags 13 via the branching portion 12g. In other words, the transfer tube 12d forms a flow path for transferring the buffy coat BC contained in the BC bag 13 to the BC separation bag 12a.

[0018] The transfer tube 12e connects the BC separation bag 12a and the PC collection bag 12c. The transfer tube 12e constitutes a flow path for transferring the platelet concentrate PC, which is the supernatant obtained by centrifuging the buffy coat BC, to the PC collection bag 12c. The leukocyte removal filter 12b is provided midway along the transfer tube 12e. The leukocyte removal filter 12b is configured to remove leukocytes remaining in the supernatant being transferred.

[0019] The BC separation bag 12a has a capacity capable of accommodating buffy coat BC collected from multiple donors. Although not particularly limited, this embodiment will be described using an example in which one BC separation bag 12a accommodates buffy coat BC collected from five donors. In this case, a bag of approximately 600 ml can be used as the BC separation bag 12a.

[0020] The PC collection bag 12c is a bag that communicates with the BC separation bag 12a via a transfer tube 12e and is configured to store the platelet concentrate PC separated in the BC separation bag 12a. The PC collection bag 12c may have a capacity of, for example, about 600 ml.

[0021] 2, the concentrate bag set 14 is used to concentrate the platelet concentrate PC to extract the concentrated platelet concentrate CPC, and is provided as a disposable product. The concentrate bag set 14 includes a PC concentrate bag 14a, a supernatant collection bag 14b, and a transfer tube 14d that connects the PC concentrate bag 14a and the supernatant collection bag 14b.

[0022] The PC concentrate bag 14a is configured to transfer the platelet concentrate PC contained in the PC collection bag 12c and to be set in a centrifuge 18 to concentrate the platelet concentrate PC. The PC concentrate bag 14a is a container formed by overlapping a pair of rectangular resin sheets and joining their peripheral edges 143. A connection tube 14c and a transfer port 14f extend from one end 141 of the PC concentrate bag 14a. Furthermore, a connecting tube 14g extends from the other end 142 opposite to the one end 141.

[0023] The connecting tube 14c and the connecting tube 14g are in communication with the interior of the PC concentrated bag 14a. When the product is provided, the ends of the connecting tube 14c and the connecting tube 14g are welded and sealed. The connecting tube 14c or the connecting tube 14g can be connected to other tubes using a sterile joining device (not shown) as needed.

[0024] 1, and serves as a flow path for transferring the platelet concentrate PC from the PC collection bag 12c. The connection tube 14c is also connected to a connecting tube 14g of another PC concentrate bag 14a, and serves as a flow path for transferring the concentrated platelet concentrate CPC to the collection bag set 16. The connecting tube 14g is connected to another PC concentrate bag 14a, and serves as a flow path for transferring the concentrated platelet concentrate CPC.

[0025] A transfer tube 14d is connected to the transfer port 14f of the PC concentrate bag 14a. The PC concentrate bag 14a and the supernatant collection bag 14b are in communication with each other via this transfer tube 14d. The transfer tube 14d constitutes a flow path that transfers the supernatant of the platelet concentrate PC, which is produced when the PC concentrate bag 14a is centrifuged, to the supernatant collection bag 14b. A clamp 34 (see FIG. 4) of the centrifuge 18, a sensor 32, a pump, and the like, which will be described later, can be attached to the transfer tube 14d.

[0026] The supernatant collection bag 14b collects the supernatant of the platelet concentrate PC. After the concentration of the platelet concentrate PC is completed, the transfer tube 14d is cut, and the supernatant collection bag 14b is separated from the PC concentrate bag 14a and collected. The PC concentrate bag 14a separated from the supernatant collection bag 14b is subjected to freezing and thawing processes in the freezer storage 20. The supernatant in the supernatant collection bag 14b is plasma and can be used as raw plasma.

[0027] The capacities of the PC concentration bag 14a and the supernatant collection bag 14b can be the same as the capacity of the PC collection bag 12c. Although not particularly limited, when buffy coats BC collected from five donors are processed as one unit, the capacities of the PC concentration bag 14a and the supernatant collection bag 14b may be approximately 300 ml. In this case, the amount of platelet concentrate PC pooled in the PC concentration bag 14a is approximately 250 ml, and the volume of concentrated platelet concentrate CPC remaining in the PC concentration bag 14a after concentration is reduced to approximately 25 ml.

[0028] 3, the collection bag set 16 is used to separate and collect platelet lysate PL from frozen and thawed concentrated platelet concentrate CPC, and is provided as a disposable product. The collection bag set 16 includes a PL separation bag 16a, a PL collection bag 16b for collecting platelet lysate PL, and a transfer tube 16d connecting the PL separation bag 16a and the PL collection bag 16b.

[0029] The PL separation bag 16a is configured to pool the concentrated platelet concentrate (CPC) contained in the multiple PC concentration bags 14a and separate the platelet lysate (PL) from the platelet membrane residue. Because the PL separation bag 16a is a bag used in freezing and thawing processes, it is preferable to use a material that will not be damaged during the freezing and thawing processes. Therefore, ethylene-vinyl acetate copolymer (EVA) resin is more preferable as the material for the PL separation bag 16a than polyvinyl alcohol (PVC) resin, which is commonly used in blood bags and the like. Furthermore, EVA resin is also suitable for the PL separation bag 16a because it has the property of being less susceptible to adsorption of plasma proteins.

[0030] The PL separation bag 16a is further provided with a connection tube 16e and a transfer port 16f. As shown in Fig. 10, the connection tube 16e is connected to the multiple PC concentration bags 14a via branching portions 16j, forming a flow path for transferring the concentrated platelet concentrate CPC from the PC concentration bags 14a to the PL separation bag 16a. As shown in Fig. 3, a transfer tube 16d is connected to the transfer port 16f. That is, the PL separation bag 16a and the PL collection bag 16b are in communication with each other via the transfer tube 16d.

[0031] The transfer tube 16d constitutes a flow path for transferring the platelet lysate PL, which is the supernatant of the concentrated platelet concentrate CPC centrifuged in the PL separation bag 16a. The transfer tube 16d is also provided with a foreign matter removal filter 16c that removes foreign matter (about 0.2 μm) such as viruses contained in the platelet lysate PL, and the platelet lysate PL from which the foreign matter has been removed by the foreign matter removal filter 16c is collected in the PL collection bag 16b.

[0032] The PL collection bag 16b collects the platelet lysate PL. The PL collection bag 16b may be provided with a sampling pouch 16h for sampling the contents.

[0033] Ordinary blood bags can be used for the PL separation bag 16a and the PL collection bag 16b. For example, when pooling the concentrated platelet concentrate CPC from 20 PC concentrate bags 14a (i.e., for 100 donors), the nominal capacity of the PL separation bag 16a and the PL collection bag 16b can be 600 ml.

[0034] 1, the centrifuge 18 is a device that is repeatedly used in the process of producing the platelet lysate PL. The centrifuge 18 includes a box-shaped device main body 22 and a rotor 30 (centrifugal unit) that is rotatably provided within the device main body 22.

[0035] The device main body 22 holds the plates collection set 12, the concentrate bag set 14, and the collection bag set 16 either internally or externally, and has the function of controlling the centrifugation of the predetermined components in each bag. The device main body 22 is equipped with a control unit 26 that receives operations during centrifugation, displays the operating status, and controls the operation of each unit. The platelet collection set 12, the concentrate bag set 14, and the collection bag set 16 are housed in an insert unit 28 and attached to a rotor 30.

[0036] As shown in FIG. 4, a sensor 32, a clamp 34, and a pump (not shown) are attached to the insert unit 28. The insert unit 28 also includes a pressing unit 33 that presses the platelet collection set 12, the concentrate bag set 14, and the collection bag set 16. The sensor 32 detects the transfer state of a predetermined component via the transfer tubes 12e, 14d, and 16d. The sensor 32 may also detect a change in the light transmittance of the transfer tubes 12e, 14d, and 16d. The sensor 32 may also detect the displacement of the pressing unit 33, which will be described later.

[0037] 1, the rotor 30 of the device body 22 is cylindrical and circumferentially divided into multiple compartments, each of which houses an insert unit 28. A rotational drive source (not shown) for rotating the rotor 30 is provided at the bottom of the rotor 30. A pressing portion 33 for pressing the platelet collection set 12, concentrate bag set 14, and collection bag set 16 attached to the insert unit 28 is provided at a portion of the rotor 30 adjacent to the inner periphery of the insert unit 28.

[0038] The insert unit 28 is configured to be freely removable from the rotor 30, allowing easy attachment of the platelet collection set 12, the concentrate bag set 14, and the recovery bag set 16. Each part of the centrifuge 18 operates under the control of a control unit 26.

[0039] 1, the freezer storage 20 is provided with a plurality of storage sections 20a. Each storage section 20a stores a plurality of PC concentrate bags 14a whose volume has been reduced by concentration. In the freezer storage 20, the concentrated platelet concentrate CPC is subjected to a freezing process.

[0040] The platelet lysate production system 10 of this embodiment is configured as described above. The platelet lysate production method of this embodiment will now be described.

[0041] The method for producing a platelet lysate is performed according to the steps shown in the flowchart of FIG. 5. First, in step S10, a medical professional such as a doctor or nurse collects whole blood. Then, in step S20, the collected blood is centrifuged in a centrifuge 18 installed in a medical facility, a blood collection vehicle, a blood center, or the like. Centrifugation separates the red blood cells, platelets, white blood cells, and plasma of the whole blood WB according to their specific gravities. As a result, the whole blood WB is separated into concentrated red blood cells, plasma, and a buffy coat BC that appears between the concentrated red blood cells and the plasma (step S30). The buffy coat BC mainly contains platelets and white blood cells.

[0042] Then, in step S300, white blood cells are removed from the buffy coat BC to collect platelet concentrate PC. First, in step S40, buffy coats BC collected from multiple donors are pooled. That is, as shown in step S41 of FIG. 6, a platelet collection set 12, multiple BC bags 13 containing buffy coats BC, and a PAS bag 11 containing platelet additive solution (PAS) are prepared.

[0043] Next, in step S42, the BC bag 13 and the PAS bag 11 are aseptically connected to the BC separation bag 12a of the platelet collection set 12. Then, the clamp 12f is opened, and the buffy coat BC and the platelet storage solution PAS are pooled into the BC separation bag 12a. By pooling the buffy coats BC from multiple donors, variation in the buffy coat BC can be reduced.

[0044] Thereafter, in step S43, the transfer tube 12d is cut to separate the BC bag 13 and the PAS bag 11 from the platelet collection set 12.

[0045] Next, in step S50 of Fig. 5, the buffy coat BC pooled in the platelet collection set 12 is centrifuged by the centrifuge 18. That is, as shown in step S51 of Fig. 7, the platelet collection set 12 is set in the centrifuge 18, and centrifugal force is applied to the BC separation bag 12a. As a result, as shown in step S52, the buffy coat BC in the BC separation bag 12a is separated into a suspension containing red blood cells and white blood cells and a supernatant liquid of plasma containing a large amount of platelets (platelet-rich plasma).

[0046] Next, as shown in step S53, the rotor 30 of the centrifuge 18 is rotated to apply centrifugal force, and the supernatant liquid from the BC separation bag 12a is transferred to the PC collection bag 12c. The supernatant liquid from the BC separation bag 12a passes through the leukocyte removal filter 12b on its way to the PC collection bag 12c via the transfer tube 12e. This removes leukocytes remaining in the supernatant liquid. As a result, the platelet concentrate PC is collected in the PC collection bag 12c.

[0047] As shown in step S54, when the sensor 32 detects leakage of red blood cells from the BC separation bag 12a, the centrifuge 18 stops the operation of the centrifugal separator. Then, in step S60, the platelet collection set 12 is removed from the centrifuge 18, and the PC collection bag 12c is separated from the BC separation bag 12a, completing the collection of the platelet concentrate PC.

[0048] Next, in step S70 of Fig. 5, the platelet concentrate PC collected in step S60 is transferred to the concentrate bag set 14. That is, as shown in step S71 of Fig. 8, the concentrate bag set 14 and the PC collection bag 12c containing the platelet concentrate PC are prepared.

[0049] Next, in step S72, the PC collection bag 12c and the PC concentrate bag 14a are aseptically connected via the connecting tube 14c, and the platelet concentrate PC in the PC collection bag 12c is transferred to the PC concentrate bag 14a.

[0050] Next, in step S73, the connecting tube 14c is cut to separate the PC collection bag 12c, thereby completing the transfer step in step S70 of FIG.

[0051] Next, in step S400, concentrated platelet concentrate CPC is prepared. First, in step S80, the platelet concentrate PC is centrifuged. That is, in step S81 of FIG. 9, the concentrate bag set 14 is set in the centrifuge 18, and the rotor 30 of the centrifuge 18 is rotated to apply centrifugal force to the PC concentrate bag 14a of the concentrate bag set 14. As a result, as shown in step S82, the platelet concentrate PC in the PC concentrate bag 14a is separated into plasma, which contains a large amount of water, and a concentrated platelet concentrate CPC (Highly Concentrated Platelet Concentrate), which contains a high concentration of platelets. The plasma is separated as the supernatant.

[0052] Next, the process proceeds to step S90 in Fig. 5 to collect plasma. That is, in step S91 in Fig. 9, while the centrifugal separator (rotor 30) is operating, the centrifuge 18 operates the pressing unit 33 to press the PC concentrated bag 14a and transfer the supernatant plasma to the supernatant collection bag 14b. Thereafter, in step S92, the centrifuge 18 detects the transfer state of the plasma to the supernatant collection bag 14b or the displacement of the pressing unit 33 with the sensor 32, and stops the operation of the centrifuge separator.

[0053] Thereafter, in step S93, the concentrate bag set 14 is removed from the centrifuge 18. Then, in step S100, the transfer tube 14d is cut to separate the supernatant collection bag 14b from the PC concentrate bag 14a. Concentrated platelets remain in the PC concentrate bag 14a, and the concentration of platelets in the PC concentrate bag 14a is complete. The volume of the concentrated platelet concentrate CPC in the PC concentrate bag 14a is reduced to about 1 / 10 of the original volume of the platelet concentrate PC. In this way, the concentration of the platelet concentrate PC in step S400 of FIG. 5 is completed.

[0054] Next, in step S500 of Fig. 5, the concentrated platelet concentrate CPC is frozen and thawed. That is, steps S110 and S130, in which the PC concentrate bag 14a is stored in the freezer storage 20 and the concentrated platelet concentrate CPC is frozen, and steps S120 and S140, in which the frozen concentrated platelet concentrate CPC is thawed, are alternately performed multiple times. In the illustrated example, the process is repeated twice, but this is not limited to this; freezing and thawing may be performed only once. Alternatively, freezing and thawing may be repeated three or more times. Performing freezing and thawing multiple times is preferable because it more reliably destroys the platelet membranes and improves the yield of platelet lysate PL.

[0055] Next, in step S600, the platelet lysate PL is collected after freezing and thawing. First, in step S150, the frozen and thawed concentrated platelet concentrate CPC is pooled. That is, as shown in step S151 of FIG. 10, a plurality of PC concentrated bags 14a and a collection bag set 16 are prepared. The PC concentrated bags 14a are connected in series by connecting the connecting tube 14g to the connecting tube 14c of other PC concentrated bags 14a. This allows a large number of PC concentrated bags 14a to be connected without increasing the number of branching sections. Furthermore, a plurality of groups of PC concentrated bags 14a connected in series are prepared.

[0056] Next, in step S152, a group of multiple PC concentrate bags 14a is connected to the collection bag set 16 via the branching portion 16j. Then, the clamp 16g is opened, and the frozen and thawed concentrated platelet concentrate CPC in the PC concentrate bags 14a is pooled into the PL separation bag 16a of the collection bag set 16. In the illustrated example, the frozen and thawed concentrated platelet concentrate CPC from 20 PC concentrate bags 14a is contained in the PL separation bag 16a. Therefore, frozen and thawed concentrated platelet concentrate CPC from 100 donors is contained in the PL separation bag 16a, thereby achieving uniform quality.

[0057] Next, in step S153, the connecting tube 16e is cut to separate the PC concentrate bag 14a from the collection bag set 16. This completes the pooling process of the concentrated platelet concentrate CPC after freezing and thawing in step S150 of FIG.

[0058] Next, in step S160 of Fig. 5, the frozen and thawed concentrated platelet concentrate CPC is centrifuged. That is, in step S161 of Fig. 11, the collection bag set 16 is set in the centrifuge 18, and the centrifuge 18 is operated to apply centrifugal force to the PL separation bag 16a. As a result, as shown in step S162, the frozen and thawed concentrated platelet concentrate CPC in the PL separation bag 16a is separated into cell membrane residue and a supernatant consisting of platelet lysate PL.

[0059] Next, in step S170 of Fig. 5, the supernatant is collected. That is, in step S171 of Fig. 11, the centrifuge 18 presses the PL separation bag 16a while applying centrifugal force to the PL separation bag 16a. As a result, the supernatant consisting of the platelet lysate PL is transferred to the PL collection bag 16b through the transfer tube 16d. At this time, the supernatant consisting of the platelet lysate PL is sterilized (including the removal of foreign matter such as viruses) by passing through the foreign matter removal filter 16c.

[0060] Thereafter, in step S172, the centrifuge 18 collects a predetermined amount of platelet lysate PL into the PL collection bag 16b, and then stops operating.

[0061] Thereafter, in step S173, the collection bag set 16 is removed from the centrifuge 18, and the PL collection bag 16b is detached to collect the platelet lysate PL. This completes the production of the platelet lysate PL of this embodiment. The PL collection bag 16b contains platelet lysate PL collected from, for example, 100 donors.

[0062] The platelet lysate production method, production system 10, and collection bag set 16 of the present embodiment described above have the following advantages.

[0063] In the method for producing platelet lysate according to the present embodiment, the platelet concentrate PC is concentrated before being frozen. This reduces the volume of the concentrated platelet concentrate CPC in the PC concentrate bag 14a that undergoes the freezing process, allowing platelets collected from multiple donors to be frozen even in a small-scale freezer 20. Furthermore, even when pooling the concentrated platelet concentrate CPC from multiple donors when collecting the platelet lysate PL, the volume does not become too large, providing excellent operability.

[0064] In the method for producing a platelet lysate, the steps of preparing concentrated platelet concentrate CPC (steps S70, S400) may include a step of transferring platelet concentrate PC collected from multiple donors to a PC concentrated bag 14a (step S70), a step of setting the PC concentrated bag 14a in a centrifuge 18 and centrifuging the platelet concentrate PC (step S80), and a step of transferring the supernatant in the PC concentrated bag 14a to a supernatant collection bag 14b and removing it (step S90). This allows the water (plasma) that occupies most of the volume of the platelet concentrate PC to be removed, and the volume of the PC concentrated bag 14a to be processed to be significantly reduced.

[0065] In the method for producing a platelet lysate, the step of collecting the platelet lysate PL (step S600) may include the steps of pooling the concentrated platelet concentrate CPC contained in multiple PC concentration bags 14a into a PL separation bag 16a (step S150), placing the PL separation bag 16a in a centrifuge 18 to separate the concentrated platelet concentrate CPC into a precipitate consisting of cell membrane residues and a supernatant containing the platelet lysate PL (step S160), and collecting the supernatant containing the platelet lysate PL into a PL collection bag 16b (step S170). Pooling the concentrated platelet concentrate CPC allows preparations using platelets from multiple donors to be produced even with small-scale equipment. This allows for consistent quality.

[0066] In the production of platelet lysate, the concentrated platelet concentrate CPC may be frozen and thawed multiple times, which allows for more effective destruction of the platelet membranes and efficient extraction of platelet lysate PL.

[0067] The platelet lysate production system 10 of this embodiment includes a concentrate bag set 14 having a PC concentrate bag 14a containing platelet concentrate PC and a supernatant collection bag 14b connected to the PC concentrate bag 14a via a transfer tube 14d, a rotor 30 that applies centrifugal force to the concentrate bag set 14, a pressing unit 33 that presses the PC concentrate bag 14a to transfer the supernatant liquid from the PC concentrate bag 14a to the supernatant collection bag 14b, and a centrifuge 18 that has a sensor 32 that detects the amount of transferred supernatant liquid. The centrifuge 18 may be configured to transfer a specified amount of supernatant liquid obtained by centrifuging the platelet concentrate PC from the PC concentrate bag 14a to the supernatant collection bag 14b and then stop the rotor 30. This allows the process of concentrating the platelet concentrate PC to be performed without manual intervention, improving productivity.

[0068] The concentrate bag set 14 of this embodiment includes a PC concentrate bag 14a provided with a connecting tube 14c, a transfer port 14f, and a linking tube 14g, a transfer tube 14d connected to the transfer port 14f, and a supernatant collection bag 14b connected to the PC concentrate bag 14a via the transfer tube 14d. By providing the linking tube 14g in addition to the connecting tube 14c, multiple PC concentrate bags 14a can be connected in series, improving the workability when collecting the platelet lysate PL.

[0069] In the concentrated bag set 14, the PC concentrated bag 14a may have a connecting tube 14c and a transfer port 14f formed at one end, and a linking tube 14g provided at the other end opposite the connecting tube 14c. With this configuration, if multiple PC concentrated bags 14a are connected in series with the connecting tubes 14c and the linking tubes 14g arranged vertically, the concentrated platelet concentrate CPC can be easily collected by gravity, improving workability.

[0070] (Second embodiment) In this embodiment, an example is described in which a step of removing plasma components and coagulation factors from concentrated platelet concentrate CPC using a cleaning solution is added to the step of concentrating platelet concentrate PC (step S400) in the method for producing platelet lysate described with reference to Figures 1 to 11.

[0071] As shown in FIG. 12, the concentrate bag set 14A of this embodiment includes a PC concentrate bag 14a, a supernatant collection bag 14b, and a washing solution bag 14h. The washing solution bag 14h contains washing solution such as platelet preservative solution (PAS) or physiological saline. The washing solution bag 14h is connected to the PC concentrate bag 14a via a washing solution tube 14i. As shown in the figure, the washing solution tube 14i is connected to the PC concentrate bag 14a at the same end as the connecting tube 14g. That is, the washing solution tube 14i is provided at the end opposite the end where the transfer tube 14d is formed. During centrifugation, the separated concentrated platelet concentrate (CPC) collects on the outer wall of the centrifuge. By introducing washing solution from the washing solution tube 14i into the PC concentrate bag 14a during centrifugation, the concentrated platelet concentrate (CPC) can be expected to be agitated with the washing solution.

[0072] In this embodiment, the supernatant collection bag 14b contains the plasma separated from the PC concentration bag 14a as well as the cleaning fluid in the cleaning fluid bag 14h. Therefore, it is preferable that the volume of the supernatant collection bag 14b be larger than the volume of the PC concentration bag 14a and the volume of the cleaning fluid contained in the cleaning fluid bag 14h.

[0073] The method for producing a platelet lysate according to this embodiment is described below. The process is the same as that shown in FIG. 8 up to the step of placing the concentrate bag set 14A in the centrifuge 18 and applying centrifugal force to the PC concentrate bag 14a to perform centrifugation. Thereafter, in step S91A of FIG. 13, in the step of recovering the separated supernatant, the cleaning solution from the cleaning solution bag 14h is poured into the PC concentrate bag 14a to wash the concentrated platelet concentrate CPC. This replaces the plasma in the concentrated platelet concentrate CPC with the cleaning solution, and coagulation factors such as fibrinogen in the concentrated platelet concentrate CPC are washed away together with the cleaning solution.

[0074] The washing solution is transferred to the supernatant collection bag 14b together with the separated plasma. The centrifuge 18 detects the completion of the transfer of the specified amount of washing solution and plasma based on the detection result of the sensor 32, and stops operation (step S92A). This completes the concentrating and washing steps of the platelet concentrate PC in the method for producing a platelet lysate of this embodiment.

[0075] The platelet lysate production method and the concentrated bag set 14A of this embodiment described above have the following advantages.

[0076] In the method for producing platelet lysate according to this embodiment, when the platelet concentrate PC in the PC concentrate bag 14a is centrifuged, a cleaning solution is supplied to the PC concentrate bag 14a to wash the platelet concentrate PC. This replaces the plasma in the platelet concentrate PC with the cleaning solution, and also removes coagulation factors such as fibrinogen. As a result, it is no longer necessary to add an anticoagulant such as heparin to the platelet lysate PL.

[0077] The concentrate bag set 14A of this embodiment further includes a washing solution bag 14h that is connected to the PC concentrate bag 14a via a washing solution tube 14i and supplies washing solution to the PC concentrate bag 14a, thereby enabling the concentration and washing of the platelet concentrate PC.

[0078] (Third embodiment) In the embodiments described with reference to Figures 1 to 13, a buffy coat BC is separated from whole blood WB collected from a donor, and a platelet concentrate PC is obtained from the buffy coat BC. However, the present invention is not limited to these embodiments. In the embodiments described below, an example is described in which a platelet concentrate PC is directly collected by component blood collection from a donor.

[0079] The method for producing a platelet lysate of this embodiment uses a bag set 50 shown in Figure 14. The bag set 50 includes a blood separation chamber 54, a reservoir 60a, a plasma bag 60b, a concentrate bag set 68, tubing 59 connecting these, a cassette 53 to which the tubing 59 configured in a predetermined path is held or connected, a blood collection section 51 equipped with a puncture needle, and an initial blood bag 52. Of these, the cassette 53 is attached to the centrifuge 18 (see Figure 1) as the centrifugal separation section 58 of the blood separation chamber 54. To prevent contamination and for sanitation, the bag set 50 is disposed of after a single use.

[0080] Whole blood WB collected from blood collection section 51 is introduced into blood separation chamber 54 after initial blood collection in initial blood bag 52. Blood separation chamber 54 is formed in a strip-shaped bag and configured to be attached to rotor 30 of centrifuge 18. Blood separation chamber 54 is a flexible bag that has a blood separation section therein to which whole blood WB from a donor is supplied, and can be easily bent. The whole blood WB introduced into the blood separation section of blood separation chamber 54 is centrifuged as it flows from one end to the other.

[0081] A leukocyte removal chamber 56 is provided near one end of the blood separation chamber 54. The leukocyte removal chamber 56 uses centrifugal force to remove leukocytes from the whole blood WB flowing within the blood separation chamber 54. A cassette 53 is connected to the blood separation chamber 54 via a tube 59a.

[0082] Tubes 59b and 59c are connected to cassette 53. Cassette 53 is connected to blood collection section 51 and initial blood flow bag 52 via tube 59b. Cassette 53 is also connected to reservoir 60a and concentrated bag set 68 via tube 59c. Tube 59c branches into tube 57a leading to reservoir 60a, tube 57b leading to plasma bag 60b, and tube 61 leading to concentrated bag set 68.

[0083] The concentrated bag set 68 includes a platelet collection bag 62, a platelet lysate bag 66, and a foreign matter removal filter 64. The platelet collection bag 62 is connected to the tube 61, and platelets separated in the blood separation chamber 54 are contained in the platelet collection bag 62. The platelet collection bag 62 is connected to the platelet lysate bag 66 via a tube 63. The tube 63 is provided with a foreign matter removal filter 64.

[0084] Whole blood WB introduced from blood collection unit 51 is introduced into blood separation chamber 54 via cassette 53. The whole blood WB is separated in blood separation chamber 54, and the separated platelets are collected in platelet collection bag 62 via tube 61. Red blood cells and plasma separated in blood separation chamber 54 are collected in reservoir 60a and plasma bag 60b, respectively. Of the separated red blood cells, the portion of red blood cells that cannot be collected in blood separation chamber 54 is collected in reservoir 60a. After centrifugation is complete, the red blood cells are returned to the donor via blood collection unit 51. The plasma collected in plasma bag 60b can be used as a pharmaceutical raw material.

[0085] Hereinafter, the method for producing a platelet lysate according to this embodiment using the bag set 50 will be described with reference to FIG.

[0086] First, blood components are collected from a donor (step S202). To collect blood components, the donor is punctured with the puncture needle of the blood collection unit 51 of the bag set 50 shown in FIG. 14 to collect blood (whole blood WB). After initial blood is removed in the initial blood bag 52, the whole blood WB is introduced into the blood separation chamber 54 of the centrifugal separator 58. In the blood separation chamber 54, the whole blood WB is separated into white blood cells, platelets, plasma, and red blood cells. Of these, the white blood cells are separated and removed in the white blood cell removal chamber 56. The plasma is collected in the plasma bag 60b via tubes 59a, 59c, and 57b. The red blood cells are collected in the reservoir 60a via tubes 59a, 59c, and 57a. The red blood cells collected in the reservoir 60a are then returned to the donor.

[0087] Meanwhile, the platelet concentrate PC is received in the platelet collection bag 62 of the concentrate bag set 68 through the tubes 59a, 59c, and 61 (step S204). In this embodiment, the platelet concentrate PC can be collected directly from the blood collector into the concentrate bag set 68. In the concentrate bag set 68, the tube 63 is initially closed with a clamp or the like, and the platelet collection bag 62 and the platelet lysate bag 66 are not in communication with each other.

[0088] Thereafter, concentrated bag set 68 is separated from bag set 50 (step S206). Concentrated bag set 68 is separated in a sterile manner by cutting tube 61 while sealing it by heat welding using a high-frequency welding device (tube sealer).

[0089] Thereafter, the separated concentrate bag set 68 is cooled in a freezer to freeze the platelet concentrate PC in the platelet collection bag 62 (step S207). Next, the concentrate bag set 68 is returned to room temperature to thaw the frozen platelet concentrate PC (step S208). The freezing and thawing process (step S210) including steps S207 and S208 may be repeated multiple times. This operation destroys the platelet membranes and produces platelet lysate PL.

[0090] Next, the platelet lysate PL is collected in the platelet lysate bag 66 (step S212). The concentrate bag set 68 is set in the centrifuge 18 (see FIG. 1), and the contents of the platelet collection bag 62 are centrifuged. The platelet lysate PL is separated as the supernatant from the platelet collection bag 62. Thereafter, the tube 63 is opened to connect the platelet collection bag 62 with the platelet lysate bag 66, and the supernatant from the platelet collection bag 62 is transferred to the platelet lysate bag 66. As the platelet lysate PL is transferred through the tube 63, it passes through a foreign matter removal filter 64, whereby foreign matter is removed. Thereafter, the platelet lysate bag 66, from which the platelet lysate PL has been collected, is aseptically separated by cutting the tube 61 with a tube sealer while heat welding the tube 61. This completes the production of the platelet lysate PL.

[0091] In addition, if the bag set 50 does not have a leukocyte removal chamber 56, as shown in Figure 16, in the component blood collection (step S202), after obtaining a buffy coat BC containing platelets and leukocytes, a step of centrifuging the buffy coat BC to remove the leukocytes can be added, thereby collecting the platelet concentrate PC in the concentrate bag set 68.

[0092] Furthermore, prior to the blood component collection by centrifugation (step S202), the whole blood WB provided by the donor may be passed through a leukocyte removal filter to remove leukocytes.

[0093] The method for producing a platelet lysate according to this embodiment has the following advantages.

[0094] The method for producing platelet lysate includes the steps of collecting platelets and other components by centrifuging the donor's whole blood (WB) to collect platelet concentrate (PC) and returning other components to the donor, freezing and thawing the platelet concentrate (PC), and, after the freezing and thawing steps, centrifuging the platelet concentrate (PC) to collect platelet lysate (PL). This reduces the burden on the donor by allowing only platelets to be collected from the donor and the other components to be returned to the donor. Furthermore, processing is simplified and fewer bags are required, reducing waste.

[0095] In the above-described method for producing a platelet lysate, when the donor's whole blood WB is centrifuged, leukocytes may be removed in the leukocyte removal chamber 56. This eliminates the need for an additional step for leukocyte removal and simplifies the configuration of the bag set 50.

[0096] (Fourth embodiment) A bag set 80 of this embodiment shown in FIG. 16 is configured so that, when used to collect whole blood WB, it can recover platelets from a leukocyte removal filter 74 that is discarded after use.

[0097] Bag set 80 includes blood collection section 51 equipped with a puncture needle, initial blood bag 52 for accommodating initial blood flow, first whole blood bag 70 for accommodating whole blood WB flowing in from blood collection section 51, second whole blood bag 76 connected downstream of first whole blood bag 70, leukocyte removal filter 74 connected between first whole blood bag 70 and second whole blood bag 76, and preservative solution bag 78 and plasma collection bag 77 connected to second whole blood bag 76. Bag set 80 further includes collection solution bag 72 connected downstream of leukocyte removal filter 74, and concentration bag set 68 connected upstream of leukocyte removal filter 74.

[0098] The first whole blood bag 70 is made of a flexible resin film or the like, and has a first port 70a and a second port 70b. The blood collection section 51 and the initial blood bag 52 are connected to the first port 70a of the first whole blood bag 70 via a tube 75a. The second port 70b of the first whole blood bag 70 is connected to a tube 75b, which is in turn connected to a second whole blood bag 76. The second port 70b is configured to be closable. A leukocyte removal filter 74 is connected to the tube 75b.

[0099] The leukocyte removal filter 74 is made of, for example, a polyester-based ultrafine microfiber nonwoven fabric and is configured to remove leukocytes from whole blood. A first valve 75c is provided in a tube 75b between the leukocyte removal filter 74 and the first whole blood bag 70, and a second valve 75d is provided in a tube 75b between the leukocyte removal filter 74 and the second whole blood bag 76. A concentrate bag set 68 is connected to the tube 75b between the first valve 75c and the leukocyte removal filter 74. A clamp 71 is provided between the concentrate bag set 68 and the tube 75b. A recovery liquid bag 72 is connected to the tube 75b between the second valve 75d and the leukocyte removal filter 74. A clamp 73 is provided between the recovery liquid bag 72 and the tube 75b.

[0100] The second whole blood bag 76 has a first port 76a and a second port 76b. The first whole blood bag 70 is connected to the first port 76a of the second whole blood bag 76 via a tube 75b. Whole blood WB from which leukocytes have been removed is introduced into the second whole blood bag 76 via the first port 76a. A tube 79 is connected to the second port 76b of the second whole blood bag 76, and a plasma collection bag 77 and a preservative solution bag 78 are connected to the second port 76b via the tube 79.

[0101] The preservative solution bag 78 contains a blood preservative solution (MAP) and is configured to be able to supply the preservative solution to the whole blood WB in the second whole blood bag 76. The plasma collection bag 77 is configured to be able to collect plasma produced when the whole blood WB in the second whole blood bag 76 is centrifuged.

[0102] Although the bag set 80 of this embodiment is configured as described above, the concentrate bag set 68, the collection solution bag 72, the plasma collection bag 77, and the preservative solution bag 78 do not need to be included from the beginning. For example, as shown in Fig. 18A, the bag set 90 may be configured without the concentrate bag set 68 and the collection solution bag 72. In this case, when leukocyte removal from the whole blood WB is completed, the concentrate bag set 68 and the collection solution bag 72 shown in Fig. 18B may be connected to the bag set 90 using a sterile connecting device.

[0103] Hereinafter, the method for producing the platelet lysate of this embodiment will be described with reference to FIG.

[0104] First, whole blood WB is collected from a donor (step S302). This step S302 is performed by puncturing the blood vessel of the donor with the blood collection unit 51, removing initial blood flow with the initial blood flow bag 52, and then storing the whole blood WB in the first whole blood bag 70, as shown in FIG.

[0105] Next, in step S304 of FIG. 17, white blood cells and platelets are trapped in the leukocyte removal filter 74. That is, the whole blood WB from the first whole blood bag 70 of FIG. 16 is transferred to the second whole blood bag 76 via the second port 70b and the tube 75b. On the way, the whole blood WB passes through the leukocyte removal filter 74, and white blood cells are trapped and removed by the leukocyte removal filter 74. Platelets are trapped in the leukocyte removal filter 74 along with the white blood cells. Thereafter, the second whole blood bag 76 is removed. The whole blood WB from the second whole blood bag 76 is used for various purposes, such as a raw material for pharmaceutical preparations or blood for transfusion. Conventionally, the leukocyte removal filter 74 would have been discarded as is, but as described above, it contains platelets that are effective as a cell culture medium. Therefore, in this embodiment, the platelets contained in the leukocyte removal filter 74 are recovered and utilized.

[0106] That is, in step S306 of FIG. 17, the platelet collection solution is caused to flow (backflow) from the outflow side to the inflow side of the whole blood WB of the leukocyte removal filter 74. The platelet collection solution may be, for example, a platelet storage solution (PAS). In step S306, in the bag set 80 of FIG. 16, the first valve 75c and the second valve 75d are closed, and the clamps 71 and 73 are opened. This places the collection solution bag 72 in communication with the downstream side of the leukocyte removal filter 74, and the concentrate bag set 68 in communication with the upstream side of the leukocyte removal filter 74. The platelet collection solution in the collection solution bag 72 is then pushed out, causing the platelet collection solution to flow (backflow) into the leukocyte removal filter 74, thereby cleaning it. The platelets collected from the leukocyte removal filter 74 are collected together with the platelet collection solution into the platelet collection bag 62 of the concentrate bag set 68.

[0107] 19, the operation of step S306 may be performed with the leukocyte removal filter 74 removed from the bag set 80. In this case, the collection fluid bag 72 is connected to the outflow (downstream) side of the leukocyte removal filter 74, and the concentrate bag set 68 is connected to the upstream side of the leukocyte removal filter 74. Then, by causing the platelet collection fluid to flow backward from the downstream side of the leukocyte removal filter 74, the platelets captured by the leukocyte removal filter 74 can be collected.

[0108] Thereafter, in step S308 of Figure 17, the concentrate bag set 68 is separated. Next, in step S310, platelets are concentrated. That is, the concentrate bag set 68 is set in the centrifuge 18 (see Figure 1) and centrifuged. As a result, the liquid containing a high concentration of platelets precipitates, and other components are separated as a supernatant. By removing the supernatant from the platelet collection bag 62, a platelet concentrate PC is obtained in the platelet collection bag 62. Note that, when centrifuging the platelet-containing liquid, it is preferable to simultaneously remove white blood cells, which are harmful to cell culture.

[0109] Next, in step S312, the platelet concentrate PC in the concentrate bag set 68 is frozen and thawed. The freezing and thawing in step S312 may be performed multiple times, thereby destroying the cell membranes of the platelets.

[0110] Next, in step S314, the platelet lysate PL is collected. That is, the concentrate bag set 68 is set in the centrifuge 18, and the contents of the platelet collection bag 62 are centrifuged. As a result, the platelet lysate PL is obtained as a supernatant in the platelet collection bag 62. The supernatant is collected in the platelet lysate bag 66, thereby obtaining the platelet lysate PL.

[0111] The method for producing a platelet lysate and the bag set 80 of this embodiment have the following advantages.

[0112] The method for producing a platelet lysate according to this embodiment includes the steps of: collecting whole blood WB from a donor (S302); passing the collected whole blood WB through a leukocyte removal filter 74 to remove leukocytes (S304); passing a platelet collection solution through the leukocyte removal filter 74 in the opposite direction to the flow of the whole blood WB to collect platelets contained in the leukocyte removal filter 74 as a platelet-containing solution (S306); further centrifuging the platelet-containing solution to remove the supernatant and concentrate the platelets to prepare a platelet concentrate PC (S310); freezing and thawing the platelet concentrate PC (S312); and centrifuging the platelet concentrate PC after the freezing and thawing step to collect the supernatant as a platelet lysate PL (S314). This allows platelets to be collected and reused from the leukocyte removal filter 74, which would otherwise be discarded when the whole blood was collected.

[0113] In the above-described method for producing a platelet lysate, a step of removing leukocytes from the platelet-containing solution may be performed simultaneously with or before the step of concentrating the platelet-containing solution (step S310), which is preferable because it allows leukocytes harmful to cell culture to be removed.

[0114] The above-described method for producing a platelet lysate may include a step of pooling platelet-containing liquid collected from multiple donors before the step (S310) of concentrating the platelet-containing liquid. That is, the platelet-containing liquid collected from multiple leukocyte removal filters 74 is collected in a platelet collection bag 62, which is then centrifuged. This reduces the number of centrifugation operations, which is advantageous in that it improves workability and production efficiency.

[0115] The bag set 80 of this embodiment includes a first whole blood bag 70, a leukocyte removal filter 74 connected downstream of the first whole blood bag 70, a second whole blood bag 76 connected downstream of the first whole blood bag 70 via the leukocyte removal filter 74, a recovery solution bag 72 connected downstream of the leukocyte removal filter 74 and containing a platelet detachment solution (recovery solution) that detaches platelets from the leukocyte removal filter 74, and a concentrate bag set 68 connected upstream of the leukocyte removal filter 74. This allows platelets adhering to the leukocyte removal filter 74, which removes leukocytes from the whole blood WB, to be recovered in the concentrate bag set 68 by causing the platelet detachment solution (recovery solution) to flow backward.

[0116] In the above bag set 80 , the concentrate bag set 68 may include a platelet collection bag 62 and a platelet lysate bag 66 connected to the platelet collection bag 62 via tubing 63 .

[0117] (Fifth embodiment) The bag set 80A of this embodiment shown in FIG. 20 is capable of collecting platelets at the same time as collecting red cell concentrate (RCC) and platelet poor plasma (PPP) by centrifuging whole blood WB.

[0118] In the bag set 80A of this embodiment, components similar to those in the bag set 80 of Fig. 16 are assigned the same reference numerals, and detailed description thereof will be omitted. As shown in the figure, in the bag set 80A, a second whole blood bag 76A is connected downstream of a first whole blood bag 70 via a leukocyte removal filter 74A. The leukocyte removal filter 74A is provided in a tube 75b connecting the first whole blood bag 70 and the second whole blood bag 76A. A collection fluid bag 72A is connected to the tube 75b upstream of the leukocyte removal filter 74A, and a residual blood collection bag 62A is connected to the tube 75b downstream of the leukocyte removal filter 74A.

[0119] The leukocyte removal filter 74A is made of, for example, a polyester-based ultrafine microfiber nonwoven fabric, and removes white blood cells (WBCs) from whole blood while allowing platelets to pass through. The collection fluid bag 72A is connected to a tube 75b via a clamp 73. The collection fluid bag 72A contains a cleaning fluid such as a platelet additive solution (PAS).

[0120] The residual blood collection bag 62A is connected to the tube 75b via a clamp 83. The residual blood collection bag 62A is a bag used to collect residual blood remaining in the leukocyte removal filter 74A after the whole blood has been transferred to the second whole blood bag 76A. The residual blood collection bag 62A is configured as a bag made of a flexible resin that can be centrifuged. A separation bag 66A may be connected to the residual blood collection bag 62A to facilitate the separation and collection of components, including platelets, that have been centrifuged. In this case, the residual blood collection bag 62A and the separation bag 66A form a collection bag set 68A.

[0121] The second whole blood bag 76A is a top-and-bottom type bag having ports (outlets) at the top and bottom, and as shown in the example shown, has a first port 76a and a second port 76b on the top side and a third port 76c on the bottom side. A tube 75b is connected to the first port 76a on the top side. A plasma collection bag 77 is connected to the second port 76b via a tube 79. The plasma collection bag 77 contains platelet poor plasma (PPP) centrifuged in the second whole blood bag 76A. An easily breakable portion is provided at the second port 76b, and is closed until just before the platelet poor plasma is transferred.

[0122] A red blood cell bag 84 is connected to the third port 76c of the second whole blood bag 76A via a tube 82. The red blood cell bag 84 contains concentrated red blood cells that have been centrifuged in the second whole blood bag 76A. An easily breakable portion is provided in the third port 76c, and is closed until just before the transfer of the concentrated red blood cells. A preservative solution bag 78 is connected to the red blood cell bag 84 via a tube 86.

[0123] A red blood cell preservation solution such as MAP solution is sealed in the preservation solution bag 78. An easily breakable section is provided at the end of the tube 86 connecting the preservation solution bag 78 and the red blood cell bag 84, and is broken and opened just before use.

[0124] Hereinafter, the method for producing the platelet lysate of this embodiment will be described with reference to FIGS.

[0125] First, whole blood WB is collected from a donor (step S401). The whole blood WB from the donor is stored in the first whole blood bag 70.

[0126] Next, the whole blood WB in the first whole blood bag 70 is transferred to the second whole blood bag 76A via the tube 75b. At this time, leukocytes in the whole blood are removed by the leukocyte removal filter 74A (step S402). After the transfer of the whole blood to the second whole blood bag 76A is completed, the second valve unit 75d is closed.

[0127] Next, residual blood in the leukocyte removal filter 74A is collected (step S403). The collection of residual blood is performed by closing the first valve unit 75c and the second valve unit 75d in Fig. 22, opening the clamps 73 and 83, and flowing the platelet preserving solution PAS (line liquid) from the collection solution bag 72A toward the residual blood collection bag 62A. The leukocyte removal filter 74A is washed with the platelet preserving solution PAS, and the resulting rinse liquid is stored in the residual blood collection bag 62A.

[0128] Thereafter, the residual blood collection bag 62A is cut off from the tube 75b at part A in Fig. 22 (step S404). The cutting from the tube 75b is performed using a sterile cutting device, and the ends are heat-sealed simultaneously, thereby cutting in a sterile manner. Also, in step S404, the tube 75b is cut on the downstream side of the leukocyte removal filter 74 (part B in Fig. 22), and the second whole blood bag 76A is cut off.

[0129] Next, the second whole blood bag 76A, the plasma collection bag 77, the red blood cell bag 84, and the preservative solution bag 78 are set in the centrifuge 18 (see FIG. 1), and the whole blood in the second whole blood bag 76A is centrifuged (step S405). By centrifugation, the whole blood in the second whole blood bag 76A is separated into platelet-poor plasma (PPP), platelet-rich plasma (PRP), and concentrated red blood cells (RCC) from the upper layer. The platelet-poor plasma (PPP) is then transferred to the plasma collection bag 77, and the concentrated red blood cells (RCC) are transferred to the red blood cell bag 84. The platelet-rich plasma (PRP) remains in the second whole blood bag 76A.

[0130] Next, the second whole blood bag 76A is subjected to a second centrifugation in order to concentrate the platelet-rich plasma (PRP) remaining in the second whole blood bag 76A into platelet concentrate (PC) (step S406).

[0131] Thereafter, the platelet lysate is produced by the processes described with reference to steps S80 to S170 in FIG.

[0132] The residual blood collection bag 62A in Fig. 22 is also centrifuged to separate the buffy coat BC from the residual blood. The buffy coat BC separated in the residual blood collection bag 62A is pooled, and a platelet lysate is produced by the process described with reference to steps S40 to S170 in Fig. 5.

[0133] As described above, in this embodiment, the leukocyte removal filter 74A passes platelets (removing only leukocytes), and the filtered whole blood is centrifuged to separate it into three layers: platelet-poor plasma (PPP), platelet-rich plasma (PRP), and concentrated red blood cells (RCC). The platelet-rich plasma (PRP) remaining in the second whole blood bag 76A as a middle layer component is processed into platelet concentrate (PC). Furthermore, the blood (whole blood) remaining in the leukocyte removal filter 74 is rinsed (washed) with platelet storage solution (PAS), and the rinse solution is collected in the residual blood collection bag 62A and centrifuged. This allows platelets to be collected without waste from both the whole blood that has passed through the leukocyte removal filter 74A and the blood remaining in the leukocyte removal filter 74A.

[0134] Although the present invention has been described above by citing preferred embodiments, it goes without saying that the present invention is not limited to the above embodiments, and various modifications are possible within the scope of the present invention. [Explanation of symbols]

[0135] 10...Platelet lysate production system 12...Platelet collection set 14... Concentration bag set 16... Collection bag set 18...Centrifuge 20...Freezer

Claims

1. A bag set used in a method for producing a platelet lysate, comprising: a step of capturing and removing white blood cells and platelets from whole blood; a step of recovering platelets from the captured white blood cells and platelets; a step of concentrating a platelet-containing solution containing the recovered platelets by centrifugation to prepare a platelet concentrate; a step of freezing and thawing the platelet concentrate; and a step of centrifuging the platelet concentrate that has been subjected to the freezing and thawing steps to recover a supernatant as a platelet lysate, a blood collection unit equipped with a puncture needle; a first whole blood bag for accommodating whole blood flowing in from the blood collection section; a second whole blood bag connected to the downstream side of the first whole blood bag via a first tube to which a valve can be attached; a leukocyte reduction filter connected to the first tube between the first whole blood bag and the second whole blood bag; a collection solution bag containing platelet collection solution, the collection solution bag being connected to the first tube between the leukocyte removal filter and the second whole blood bag via a third tube to which a clamp can be attached; a platelet collection bag connected to the first tube between the first whole blood bag and the leukocyte removal filter via a fourth tube to which a clamp can be attached; a platelet lysate bag connected to the platelet collection bag via a second tube that can be fitted with a valve.

2. 2. The bag set according to claim 1, further comprising: a first valve unit provided in the first tube between the leukocyte removal filter and the first whole blood bag; and a second valve unit provided in the first tube between the leukocyte removal filter and the second whole blood bag.

3. 3. The bag set according to claim 1, wherein the second tube connecting the platelet collection bag and the platelet lysate bag is provided with a foreign matter removal filter for removing foreign matter.

4. a leukocyte removal filter connected to the first tube between the first whole blood bag and the second whole blood bag; a collection solution bag containing a platelet collection solution, the collection solution bag being connected to the first tube between the leukocyte removal filter and the second whole blood bag via a third tube to which a clamp can be attached; a platelet collection bag connected to the first tube between the first whole blood bag and the leukocyte removal filter via a fourth tube to which a clamp can be attached; and a platelet lysate bag connected to the platelet collection bag via a second tube to which a valve can be attached, storing the whole blood collected from the blood collection section in the first whole blood bag; transferring the whole blood stored in the first whole blood bag to the second whole blood bag through the first tube while the collection solution bag and the platelet collection bag are fluidly separated from the first tube; a step of collecting platelet-containing fluid containing platelets adsorbed to the leukocyte removal filter into the platelet collection bag by flowing the platelet collection fluid from the collection fluid bag toward the platelet collection bag while the first whole blood bag and the second whole blood bag are fluidly separated from the leukocyte removal filter, the collection fluid bag, and the platelet collection bag; disconnecting the platelet collection bag and the platelet lysate bag from the first tube; a step of concentrating the platelet-containing solution contained in the platelet collection bag by centrifugation to prepare a platelet concentrate; freezing and thawing the platelet concentrate in the platelet collection bag; a step of centrifuging the platelet concentrate that has been subjected to the freezing and thawing steps, and recovering the supernatant as a platelet lysate in the platelet lysate bag through the second tube; A method for producing a platelet lysate, comprising:

5. 5. The method for producing a platelet lysate according to claim 4, wherein a step of removing leukocytes from the platelet-containing liquid is carried out simultaneously with or before the step of concentrating the platelet-containing liquid.

6. 5. The method for producing a platelet lysate according to claim 4, further comprising the step of pooling the platelet-containing liquid collected from a plurality of the leukocyte removal filters into the platelet collection bag before the step of concentrating the platelet-containing liquid.

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