JP2003052814A - Platelet capture filter and platelet factor collecting device - Google Patents

Abstract

PROBLEM TO BE SOLVED: To provide a platelet capture filter having a high capture rate for platelets and a low capture rate for white corpuscle in filtering a liquid to be processed containing platelets like whole blood, and to provide a platelet factor collecting device capable of collecting the platelet factors from the platelets. SOLUTION: The platelet capture filter 7 shown in Fig. 2 has a function of capturing platelets from the whole blood, and the filter is constructed by a housing 8, a filter 9 set in the housing 8, and support members 10a, 10b for the filter 9. The filter member 91 and pre-filter member 92 are respectively formed by stacking one or more disc-like porous membranes (membrane-like porous body) 911, 921. the surface of the filter member 91 is coated with a copolymer composed mainly of alkoxyalkyl(metha)acrylate and alkyl aminoalkyl(metha)acrylate, whereby the ζ-potential is set to 26 to 38 mV.

Description

Detailed Description of the Invention

[0001]

TECHNICAL FIELD The present invention relates to a platelet trapping filter and a platelet factor recovery device.

[0002]

2. Description of the Related Art Conventionally, when collecting factors derived from platelets, platelets are activated by adding platelet activating agents such as thrombin to the platelets after separating the platelets from whole blood by centrifugation. By doing so, the factor released from platelets (hereinafter referred to as “platelet factor”) is collected.

The platelet factor is, for example, Platelet derived growth factor (PDG).
F), transforming growth factor (Transforming gr
owthfactor; TGF), epidermal growth factor
owth factor (EGF), insulin-like growth factor (Insulin
like growth factor; IGF).

The collected platelet factors are used as a raw material for a wound healing promoter.

Conventionally, when an operation for activating platelets is performed, a centrifugal separation operation is performed prior to the operation.
Platelet-rich plasma (PRP), platelet-rich (PC), etc. with platelets suspended in plasma (platelet suspension)
In addition, the pretreatment (pretreatment) of washing and removing the supernatant plasma is carried out after the platelet suspension is settled in the platelet suspension, but this pretreatment requires a great deal of labor and time. There was a problem in that

As a solution to this problem, Japanese Patent Publication No. 7-507558 discloses that a platelet suspension separated by a centrifugation method is filtered through a filter, and the platelets captured by the filter are thrombin or the like. The method of recovering platelet factors by activating the platelets by adding the platelet activating agent is described.

[0007] This method can recover platelet factors in a relatively short time, but the filter used in this method is originally for mainly capturing white blood cells and white blood cells and platelets. Therefore, particularly when whole blood is filtered, a large amount of white blood cells are captured on the filter together with platelets.

Therefore, the captured white blood cells narrow the channel (pore) of the filter over time, and the narrowing of the channel causes the platelets captured by the filter to be stimulated and activated. I will end up.

As a result, the platelet factor released from the platelets passes through the filter together with the filtrate, and there is a problem that the recovery rate of the platelet factor decreases.

[0010]

DISCLOSURE OF THE INVENTION An object of the present invention is to obtain platelets having a high platelet capture rate and a low leukocyte capture rate when a liquid containing platelets such as whole blood is filtered. Another object of the present invention is to provide a trapping filter and a platelet factor recovery device that can recover platelet factors from platelets easily and in a short time.

[0011]

The above objects are achieved by the present invention described in (1) to (29) below.

(1) A platelet trapping filter for trapping platelets from a liquid to be treated containing platelets, which is made of a porous material in a housing having an inlet and an outlet, and has a zeta potential on its surface. A platelet capture filter comprising a filter containing a filter member of 26 to 38 mV.

(2) The surface of the filter member is coated with a copolymer mainly containing an alkoxyalkyl (meth) acrylate and an alkylaminoalkyl (meth) acrylate, whereby the zeta potential of the surface is 26. The platelet capture filter according to (1) above, which has a voltage of ~ 38 mV.

(3) A platelet trapping filter for trapping platelets from a liquid to be treated containing platelets, which is formed of a porous body in a housing having an inlet and an outlet, and has an alkoxyalkyl ( A platelet trapping filter comprising a filter containing a filter member coated with a copolymer mainly containing (meth) acrylate and alkylaminoalkyl (meth) acrylate.

(4) The alkoxyalkyl (meth)
The platelet capture filter according to (2) or (3) above, wherein the composition ratio of the acrylate and the alkylaminoalkyl (meth) acrylate is 85:15 to 50:50.

(5) The platelet-capturing filter according to any one of (1) to (4) above, wherein the filter member has an average pore size of 5 to 20 μm.

(6) The platelet-capturing filter according to any one of (1) to (5) above, wherein the filter member is composed of a membrane-like porous body and is formed by laminating a plurality of them.

(7) The platelet-capturing filter according to (6), wherein each of the plurality of membranous porous bodies constituting the filter member is made of a nonwoven fabric or a foam.

(8) The filter includes a pre-filter member whose average pore diameter or surface zeta potential is different from that of the filter member, and the pre-filter member is more than the filter member in the housing. The platelet capture filter according to any one of (1) to (7) above, which is located on the inlet side.

(9) The platelet filter according to (8), wherein the prefilter member has an average pore size of 30 to 100 μm.

(10) The prefilter member has a surface zeta potential of 0 to 25 mV.
The platelet capture filter described in 1.

(11) The platelet-capturing filter according to any one of (8) to (10), wherein the pre-filter member is composed of a film-like porous body and is formed by laminating a plurality of these.

(12) The platelet trapping filter according to (11), wherein each of the plurality of membranous porous bodies constituting the prefilter member is made of a nonwoven fabric or a foam.

(13) The platelet-capturing filter according to any one of (1) to (12), wherein the volume ratio of the filter member to the entire filter is 50% or more.

(14) A first line which is connected to the platelet capturing filter according to any one of (1) to (13) above and which is connected to the inlet and which supplies the liquid to be treated into the platelet capturing filter. And connected to the outlet,
It has a second line for collecting the liquid that has passed through the platelet capturing filter, and a platelet factor recovery means for activating the platelets captured by the platelet capturing filter and recovering the platelet factor released from the platelets. A platelet factor recovery device characterized by the above.

(15) The platelet factor recovery device according to (14), wherein the platelet recovery means activates the platelets by physical stimulation.

(16) The platelet factor recovery means is configured to share a part of the first line and / or the second line, and the above (14) or (1)
The platelet factor recovery device according to 5).

(17) The platelet factor recovery means has a pair of containers installed via the platelet capture filter, of which the first container is connected to the first line and the second container is connected to the second line. 7. The platelet factor recovery device according to any of (14) to (16) above, wherein the container is connected to the second line.

(18) In the middle of the first line,
The platelet factor recovery device according to (17) above, wherein a first flow path switching device is installed, and the first container is connected to the first line via the first flow path switching device. .

(19) In the middle of the second line,
The second flow path switching device is installed, and the second container is connected to the second line via the second flow path switching device. (17) or (18) above. Platelet factor recovery device.

(20) By moving a liquid for stimulating platelets between the first container and the second container, the liquid is brought into contact with the platelets captured by the platelet capture filter, The platelet factor recovery device according to any one of (17) to (19), wherein the platelet factor is activated by:

(21) The platelet factor recovery device according to the above (20), wherein the liquid is moved at least once.

(22) The platelet factor recovery device according to the above (20) or (21), wherein the flow rate of the liquid during the platelet activation operation is 0.5 to 10 mL / sec.

(23) The first container and the second container
Each of the containers is a syringe, and the platelet factor recovery device according to any one of (17) to (22) above.

(24) In the above (1), a third line for supplying a washing solution is provided in the platelet capturing filter.
The platelet factor recovery device according to any one of 4) to (23).

(25) The third line is connected to the upstream side of the platelet factor recovery means (24).
The platelet factor recovery device according to 1.

(26) The third line is the first line.
The platelet factor recovery device according to (24) or (25), which is connected in the middle of the line.

(27) The platelet factor recovery device according to any one of (24) to (26), which has a fourth line for recovering the washing liquid after passing through the inside of the platelet capture filter.

(28) The fourth line is the second line.
The platelet factor recovery device according to (27), which is connected in the middle of the line.

(29) The platelet factor recovery device according to any one of (14) to (28), wherein the liquid to be treated is whole blood.

[0041]

BEST MODE FOR CARRYING OUT THE INVENTION Hereinafter, a platelet capturing filter and a platelet factor recovery device of the present invention will be described in detail with reference to the preferred embodiments shown in the accompanying drawings.

FIG. 1 is a schematic view showing the overall structure of the platelet factor recovery device of the present invention, and FIG. 2 is an exploded perspective view (partially shown in section) showing the structure of the platelet capture filter of the present invention. .

The platelet factor recovery device 1 shown in FIG. 1 captures platelets from a liquid to be treated containing platelets, and platelet factors released from the platelets (hereinafter simply referred to as “platelet factor”).
Say ) Is a device for collecting, and a platelet capture filter 7 of the present invention, a first line (processing liquid supply line) 2, a third line (washing liquid supply line) 3,
Second line (filtered liquid recovery line) 4, fourth line (washed liquid recovery line) 5, and platelet factor recovery means 6
And have. Hereinafter, the configuration of each unit will be sequentially described.

In the following description, the case where whole blood is treated as a liquid to be treated containing platelets will be described as a representative example.

First, the platelet trapping filter 7 of the present invention will be described. As a result of intensive studies, the present inventor has found that the adsorption rate of platelets to the filter is more easily affected by the zeta potential on the surface of the filter than that of white blood cells. That is, the present inventors have found that the platelets can be more selectively captured by the filter by appropriately setting the zeta potential on the surface of the filter. The present invention has been made based on such findings.

The platelet capture filter 7 shown in FIG. 2 has a function of capturing platelets from whole blood.
It is composed of a housing 8, a filter 9 installed in the housing 8, and support members 10a and 10b of the filter 9. In other words, the platelet capture filter 7
Inside the housing 8, the filter 9 and the support member 1
It contains 0a and 10b. In the following description, in FIG. 2, the upper side is referred to as “upper” and the lower side is referred to as “lower”.

The housing 8 has a bottomed cylindrical first housing member (lower housing member) 81 and a top plate portion (ceiling) mounted (fixed) on the first housing member 81.
And a substantially cylindrical second housing member (upper housing member) 82.

At the bottom of the first housing member 81, an outflow port (liquid discharge port) 811 communicating with the inside of the housing 8 is formed to project.

At the inner lower portion of the first housing member 81, a reduced diameter portion 813 is formed by reducing the diameter of the inner cavity portion 812. The second housing member 82 is attached to the first housing member 81, and the filter 9 and the support member 1 are provided inside.
In the state in which 0a and 10b are stored (hereinafter, this state is referred to as “the assembled state of the platelet capture filter 7”), the support member 10a is installed (stored) in the reduced diameter portion 813.

In the assembled state of the platelet trapping filter 7, the filter 9 is installed (stored) in the space defined by the inner cavity 812 and the inner cavity 822 of the second housing member 82 described later. It

A fitting portion 814 having a substantially cylindrical shape is formed along the outer periphery of the upper portion of the first housing member 81. In the assembled state of the platelet capture filter 7, the fitting portion 824 of the second housing member 82 is inserted inside the fitting portion 814.

On the top plate portion of the second housing member 82,
Inflow port (liquid introduction port) 821 communicating with the inside of the housing 8
Are formed to project.

A diameter-reduced portion 823 obtained by reducing the diameter of the inner cavity portion 822 is formed in the upper portion on the inner side of the second housing member 82. In the assembled state of the platelet capture filter 7, the support member 10b is installed on the reduced diameter portion 823.

Also. A fitting portion 824 having a substantially cylindrical shape is formed along the inner periphery of the lower portion of the second housing member 82. In the assembled state of the platelet capture filter 7, the fitting portion 824 is inserted inside the fitting portion 814, and by fitting them, the first housing member 81 and the second housing member 82 are fixed. It

As the constituent material of the housing 8, for example, polyvinyl chloride, polyethylene, polypropylene,
Polystyrene, poly- (4-methylpentene-1), polycarbonate, acrylic resin, acrylonitrile-butadiene-styrene copolymer, polyester such as polyethylene terephthalate, butadiene-styrene copolymer, polyamide (for example, nylon 6, nylon 6)・
6, various resins such as nylon 6, 10 and nylon 12).

The filter 9 has a filter member 91 and a pre-filter member 92 each made of a porous material.

The filter member 91 is a main part for trapping platelets, and is formed by laminating a plurality of substantially disk-shaped porous membranes (membrane-like porous bodies) 911.

The porous membranes 911 are, for example, non-woven fabrics made of synthetic resin such as polyurethane, polyvinyl alcohol, polypropylene, polyether polyamide, polyethylene terephthalate, polybutylene terephthalate, foams (sponge-like porous materials having communicating holes). Examples of the material include densities), woven fabrics, meshes, and the like. Among these, it is particularly preferable to use a nonwoven fabric or a foam. By using a non-woven fabric or a foam made of the synthetic resin as each porous film 911, it is easy to control the pore size and the like, and there is an advantage that the productivity is excellent.

The average thickness of each porous film 911 is appropriately set depending on its constituent material, form, etc., and is not particularly limited, but for example, it is preferably about 0.05 to 5 mm, and about 0.1 to 3 mm. Is more preferable.

The number of porous membranes 911 installed is as follows.
The average thickness is appropriately set and is not particularly limited, but for example, it is preferably about 2 to 100 sheets, and 3 to 80.
It is preferable that the number is about one.

Filter member 91 (each porous film 911)
Has a surface zeta potential of, for example, 26 to 38 mV.
It is preferable that the voltage level is approximately the same, and it is more preferable that the voltage level is approximately 29 to 35 mV. By setting the zeta potential on the surface of the filter member 91 within the above range, the platelet capture filter 7 can improve the platelet capture rate and reduce the white blood cell capture rate.

Therefore, in the platelet capture filter 7,
Since platelets can be captured more selectively, the narrowing of the flow path (pore) of the filter member 91 (hereinafter, simply referred to as “clogging”) due to the increase in the amount of captured white blood cells. It can be suitably prevented (suppressed). As a result, in the platelet capture filter 7, the filter member 91
It is possible to suitably prevent (suppress) the platelets captured by the plate from being stimulated and activated by this clogging.

If the zeta potential on the surface of the filter member 91 is too low, the platelet capture rate may not be sufficient depending on the average pore size of the filter member 91, while the zeta potential on the surface of the filter member 91 may be insufficient. If is too high, the platelets may be activated when they are captured.

Examples of the method of adjusting the zeta potential on the surface of the filter member 91 include the following methods.

The surface of the filter member 91 is post-treated (eg, coated) with a substance capable of adjusting the zeta potential (eg, a substance having a cationic functional group such as an amino group).
how to.

A method in which a material having a zeta potential within the above range is used as a constituent material of the filter member 91.

In the constituent material of the filter member 91,
A method in which a substance capable of adjusting the zeta potential is contained or bound in advance and the filter member 91 is formed using such a material.

Among these, the method for adjusting the zeta potential on the surface of the filter member 91 is as follows.
It is preferable to use a method of coating the surface of No. 1 with a substance whose zeta potential can be adjusted. According to this method, the range of selection of the constituent material of the filter member 91 can be widened, and the zeta potential of the surface of the filter member 91 can be
It can be adjusted more easily and reliably.

As the substance whose zeta potential can be adjusted, any substance can be used as long as the zeta potential of the surface of the filter member 91 can be adjusted within the above range. For example, alkoxyalkyl (meth). It is preferable to use a copolymer mainly composed of an acrylate and an alkylaminoalkyl (meth) acrylate having a basic functional group. The zeta-potential of the surface of the filter member 91 can be easily adjusted by appropriately setting the amount of adhesion of the copolymer and the composition ratio of the monomers constituting the copolymer.

Examples of the alkoxyalkyl (meth) acrylate include 2-methoxyethyl acrylate (MEA) and 2-methoxyethyl methacrylate (ME).
MA) etc. can be used.

On the other hand, as alkylaminoalkyl (meth) acrylate, for example, 2- (N, N-dimethylamino) ethyl methacrylate (DMAEMA), 2-
(N, N-dimethylamino) ethyl acrylate (DM
AEA), 2- (N, N-dimethylamino) propyl acrylate (DMAPA), 2- (N, N-dimethylamino) propyl methacrylate (DMAPMA) and the like can be used.

By mainly using the filter member 91 coated with the copolymer composed of these monomers, the platelet capture filter 7 can further improve the platelet capture rate and the white blood cell capture rate. Can be reduced more reliably.

The composition ratio of the alkoxyalkyl (meth) acrylate and the alkylaminoalkyl (meth) acrylate is not particularly limited, but is, for example, 85: 1.
It is preferably about 5 to 50:50, and about 80:20.
It is more preferably about 60:40. By setting the composition ratio of each monomer within the above range, the platelet capture filter 7 can achieve both an excellent platelet capture rate and an excellent inhibitory effect on the activation of the platelets at the time of capturing the platelets. .

When the composition ratio of each monomer is less than the lower limit value, the trapping rate of platelets of the filter member 91 may be increased depending on the kind of alkoxyalkyl (meth) acrylate and / or alkylaminoalkyl (meth) acrylate. In some cases, it may not be possible to achieve a sufficient amount. On the other hand, when the composition ratio of these exceeds the upper limit value, depending on the type of alkoxyalkyl (meth) acrylate and / or alkylaminoalkyl (meth) acrylate, etc. In some cases, the effect of suppressing the activation of platelets at the time of capturing the platelets may not be sufficient.

Further, the filter member 91 (each porous film 9
The average pore diameter of 11) is not particularly limited, but for example, preferably about 5 to 20 μm, and 8 to 15 μm.
More preferably, it is about m. When the average pore size of the filter member 91 is less than the lower limit value, the filter member 91
Leukocytes may be non-specifically trapped on the surface on the inflow port 821 side (upper surface in FIG. 2), and as a result, the captured platelets are stimulated and activated due to clogging. Sometimes. On the other hand, when the average pore size of the filter member 91 exceeds the upper limit value, the filter member 91 may have a low platelet capture rate.

In other words, by setting the average pore diameter of the filter member 91 within the above range, the filter member 91
Then, it is possible to preferably prevent (suppress) increase (increase) of non-specific capture of leukocytes and decrease of capture rate of platelets.

A filter member 91 is provided in the housing 8.
A pre-filter member 92 is installed closer to the inflow port 821.

The pre-filter member 92 has a function of preventing (suppressing) clogging mainly by white blood cells in the flow path (pore) of the filter member 91.
A plurality of porous films (membrane-like porous bodies) 921 are laminated.

Each of the porous membranes 921 is made of a nonwoven fabric or a foam made of the same synthetic resin as that of the porous membrane 911, and its shape, dimensions, etc. are the same as those of the porous membrane 91.
It can be similar to 1.

The number of porous membranes 921 installed is
The thickness is appropriately set depending on the average thickness and is not particularly limited, but for example, it is preferably about 1 to 30 sheets, and preferably about 2 to 10 sheets.

As the pre-filter member 92 (each porous membrane 921), it is preferable to use one having a different average pore diameter and surface zeta potential from the filter member 91.

When the average pore diameters are different, the average pore diameter of the pre-filter member 92 is
The average pore diameter is set to be larger than 1, and specifically, 30 to
The thickness is preferably about 100 μm, more preferably about 50 to 80 μm.

On the other hand, when the surface zeta potentials are different, the surface zeta potential of the pre-filter member 92 is set to be smaller than the surface zeta potential of the filter member 91, specifically, 0 to 25 mV. It is preferably about 10 to 15 mV, more preferably about 5 to 15 mV.

With such a structure, the pre-filter member 92 can more reliably prevent (suppress) the clogging of the filter member 91.

As a method for adjusting the zeta potential on the surface of the pre-filter member 92, the same method as that described for the filter member 91 can be used.

The pre-filter member 92 (each porous membrane 921) is different from the filter member 91 in terms of either the average pore size or the surface zeta potential. May be.

In such a filter 9, the volume ratio of the filter member 91 to the entire filter is, for example,
It is preferably about 50% or more, and more preferably about 70% or more. Within such a volume ratio range, the filter member 91 and the pre-filter member 92 can exert their respective functions without any limitation.

Support members 10a and 10b are attached to the lower and upper portions of the filter 9, respectively, and are housed in the housing 8 in this state. Support members 10a, 10b
Has a function as a supporting member of the filter 9, a liquid supply to the filter 9, and a filter 9 respectively.
It has a function of smoothly discharging the liquid from the. That is, the support members 10a and 10b can also be referred to as flow channel securing members, respectively.

The supporting members 10a and 10b are composed of a ring 101 made of a flexible material such as silicone, and a mesh (diffusing member) 102 inserted inside the ring 101. There is.
The ring 101 has a thickness of, for example, about 0.1 to 2 mm.

By installing the support member 10b, a space communicating with the inflow port 821 is formed inside the ring 101, a flow path of the liquid flowing from the inflow port 821 to the filter 9 is secured, and the support member 10a is installed. Thus, a space communicating with the outflow port 811 is formed inside the ring 101, and a flow path for the liquid discharged from the filter 9 to the outflow port 811 is secured. That is, the filter 9 is
It is installed in the housing 8 so as to define a space communicating with the inflow port 821 and a space communicating with the outflow port 811.

In the support members 10a and 10b,
By installing the mesh 102 inside the ring 101, when the liquid passes through the inner space of the ring 101,
It is uniformly dispersed (diffused), and the efficiency of filtration, platelet capture, washing, etc. can be improved.

As the support members 10a and 10b,
The structure is not limited to the above, and may be, for example, a porous body made of various ceramic materials or the like.

Also, these support members 10a, 10b
Is required (for example, the outlet 81 of the housing 8).
1 and the size of the inner diameter of the inlet 821).

As described above, the platelet capture filter 7 of the present invention has an extremely high platelet capture rate and a very low white blood cell capture rate. Therefore, the platelet-capturing filter 7 of the present invention has an advantage (feature) that whole blood can be directly processed without pretreatment such as centrifugation.

Next, the whole platelet factor recovery device 1 incorporating the platelet capture filter 7 will be described.

The first line 2 is a line for supplying whole blood (liquid to be treated) into the platelet trapping filter 7, and is mainly a tube 21 and a needle tube 22 connected to one end of the tube 21. It consists of and. The other end of the tube 21 is connected to the inflow port 821 of the platelet capture filter 7 (second housing member 82).

The needle tube 22 is composed of a bottle needle, and a blood bag (not shown) in which whole blood (process liquid) is stored.
The rubber plug (plug) of can be pierced. By piercing the rubber plug with the needle tube 22, the inside of the platelet capturing filter 7 and the inside of the blood bag (container) are communicated with each other, and whole blood can be supplied into the platelet capturing filter 7.

In the middle of the tube 21, a clamp 24 is installed as a flow path opening / closing means for blocking / opening the internal flow path of the tube 21.

In the middle of the tube 21, a first three-way stopcock (first flow path switching device) 62 for connecting a syringe 61 constituting the platelet factor recovery means 6 described later near the platelet capture filter 7 is provided. Is installed.

In this embodiment, the needle tube 22 is connected to one end of the tube 21, but instead of this,
The blood bag may be directly connected.

The needle tube 22 may be composed of a puncture needle such as a winged needle. In this case, the blood can be directly supplied from the blood donor into the platelet capture filter 7 by puncturing the blood vessel of the blood donor (donor) with the puncture needle. In this case, it is preferable to connect an anticoagulant supply line for adding the anticoagulant to the blood sample in the middle of the first line.

The third line 3 is a line for supplying the washing liquid into the platelet trapping filter 7, and mainly includes the tube 31 and the needle tube 3 connected to one end of the tube 31.
It is composed of 2 and. The other end of the tube 31 is connected to the middle of the tube 21 via a Y-shaped branch connector 33. That is, the third line 3 is connected in the middle of the first line 2.

The needle tube 32 is composed of a bottle needle and can penetrate a rubber stopper (plug) of a cleaning liquid bag (not shown) containing a cleaning liquid. By piercing the rubber plug with the needle tube 32, the inside of the platelet capture filter 7 and the inside of the washing solution bag (container) can be communicated with each other to supply the washing solution into the platelet capture filter 7.

In the middle of the tube 31, this tube 3
A clamp 34 is installed as a flow path opening / closing means capable of blocking / opening the first internal flow path.

The branch connector 33 is installed upstream of the first three-way stopcock 62 of the platelet factor recovery means 6 described later. That is, the third line 3 is connected to the upstream side of the platelet factor recovery means 6.

Examples of the washing solution supplied through the third line 3 include various electrolyte solutions such as physiological saline, Hanks balanced salt solution, Dulbecco balanced salt solution, sugar solution such as glucose solution, and ACD. -A liquid, CPDA-1
A blood preservation solution such as a solution is preferably used.

In the present embodiment, the needle tube 32 is connected to one end of the tube 31, but instead of this,
The cleaning solution bag may be directly connected.

The other end of the tube 31 may be directly connected to the platelet capture filter 7.

Further, the third line 3 can be omitted if necessary. In this case, the first line 2
The needle tube 21 may be connected to the cleaning solution bag and used.

The second line 4 collects whole blood (liquid to be treated: liquid) that has passed through the platelet trapping filter 7, that is, whole blood (treated liquid) after platelets have been trapped. The line 8 is for connecting the outlet 8 of the platelet capture filter 7 (first housing member 81).
The tube 41 is connected to 11 and has the other end connected to communicate with the inside of the treated liquid recovery bag 40.

In the middle of the tube 41, a clamp 44 is installed as a flow path opening / closing means for blocking / opening the internal flow path of the tube 41.

In the middle of the tube 41, a second three-way stopcock (second flow path switching device) 64 for connecting a syringe 63 constituting the platelet factor recovery means 6 described later near the platelet capture filter 7 is provided. Is installed.

In the present embodiment, the treated liquid collecting bag 40 is connected to the other end of the tube 41, but instead of this, for example, a blood returning line and a blood returning pump, etc. Means may be provided to allow the treated liquid to be directly returned to the donor.

The fourth line 5 is a line for collecting the washing liquid (washed liquid) after passing through the inside of the platelet-capturing filter 7, and one end of which is a Y-shaped line installed in the middle of the tube 41. The tube 51 is connected to a branch connector 53 having a shape, and the other end is connected to communicate with the inside of the washed liquid recovery bag 50. That is, the fourth line 5 is connected in the middle of the second line 4.

In the middle of the tube 51, the tube 5
A clamp 54 is installed as a flow path opening / closing means capable of blocking / opening the first internal flow path.

Further, the branch connector 53 is installed on the downstream side of the second three-way stopcock 64 of the platelet factor recovery means 6 described later.

[0117] One end of the tube 51 may be directly connected to the platelet capture filter 7.

Further, the fourth line 5 is, if necessary,
It can be omitted. In this case, the cleaned liquid may be collected using the second line 4.

The platelet factor recovery means 6 has a function of activating the platelets captured by the platelet capture filter 7 and recovering the platelet factor released from the platelets. It is composed of a pair of syringes (first container and second container) 61 and 63 which are installed via the above.

Of these, the syringe (first container) 61
Is connected in the middle of the tube 21 (first line 2) via the first three-way stopcock (first flow path switching device) 62, and the syringe (second container) 63 is the second Via the three-way stopcock (second flow path switching device) 64 of the tube 4
1 (second line 4).

In the platelet factor recovery means 6, by switching the flow paths of the first three-way stopcock 62 and the second three-way stopcock 64,
The liquid can be alternately moved between the syringe 61 and the syringe 63. That is, the platelet factor recovery means 6 has a flow path (liquid circulation means) through which the liquid can move, and in this embodiment, this flow path is part of the first line 2 and the second line 4. Are configured to share.
As a result, the platelet factor recovery device 1 can be simplified and downsized.

Further, in the present embodiment, since the third line 3 is connected to the upstream side of this flow channel, as will be described later, the flow channel is washed with the washing liquid prior to the platelet factor recovery operation. Will be done. Therefore, it is possible to further reduce the mixing of unnecessary components (in particular, white blood cells) and the like into the obtained platelet factor.

The platelet factor recovery means 6 may be configured so as to share a part thereof with either the first line 2 or the second line 4, or the first line 2 or the second line 4.
The line 2 and the second line 4 may be provided separately and independently.

In the platelet factor recovery means 6 as described above, the liquid for stimulating the platelets is moved between the syringes 61 and 63 to bring them into contact with the platelets captured by the platelet capture filter 7, Activates platelets. That is, the platelet factor recovery means 6 is
It is configured to activate platelets by physical stimulation.

At this time, the platelet factor recovery means 6
Then, the platelet factor released from the platelets is collected in the syringe 61 or the syringe 63 together with the liquid.

[0126] Examples of the method for activating platelets include
Although it can be carried out by contacting an inducing substance such as thrombin with platelets, it is preferable to activate the platelets by physical stimulation. This has the advantage of being able to more reliably avoid the risk of bacteria, viruses, etc. being mixed into the obtained platelet factor when activating platelets, as compared with the case of using a biogenic inducer.

The liquid to be used is not particularly limited as long as it does not adversely affect the platelet factor and the living body, and examples thereof include physiological saline and phosphate buffer. , HEPES buffer, etc. can be used.

The flow rate of the liquid during the platelet activation operation is not particularly limited, but is, for example, 0.5 to 10
It is preferably about mL / sec, more preferably about 1 to 5 mL / sec. This makes it possible to more reliably activate the platelets.

The physical stimulus used to activate platelets includes not only the stimulus caused by the contact of the liquid with the platelets, but also various stimuli such as ultrasonic waves, electricity, and temperature change. One kind or a combination of two or more kinds can be used. When various stimuli such as ultrasonic waves, electricity, and temperature change are used as physical stimuli used for activating platelets, these various stimuli are applied to the platelets (platelet capture filter 7) by the platelet factor recovery means 6. Various means that can be provided may be provided.

The constituent material of each of the tubes 21, 31, 41 and 51 is not particularly limited, but for example, polyvinyl chloride is preferably used. This is because if these tubes are made of polyvinyl chloride, sufficient flexibility and flexibility can be obtained, so that they are easy to handle, and they are also suitable for blockage due to clamps and the like.

As the constituent materials of the branch connectors 33 and 53, the same constituent materials as those mentioned for the tube can be used.

As the constituent materials of the treated liquid recovery bag 40 and the washed liquid recovery bag 50, for example, soft polyvinyl chloride is preferably used.

Next, an example of how to use the platelet factor recovery device 1 will be described. [1] First, as shown in FIG. 1, the first line 2 and the third line 3 are respectively the platelet capture filter 7
The third line 4 and the fourth line 5 are located vertically above and are installed vertically below the platelet capture filter 7.

Further, in the portions of the first three-way stopcock 62 and the second three-way stopcock 64, the tubes 2 are respectively provided.
The internal channels 1 and 41 are in communication with each other.

At this time, the clamps 24, 34,
44 and 54 are each in a closed state, and the tubes 21, 31, 41 and 51 are
Each of the internal channels is closed.

Next, in this state, the first line 2 is inserted into the outlet of the blood bag (not shown) in which the whole blood is stored.
Needle tube 22 of the third line 3 to the rubber stopper of the cleaning liquid storage bag (not shown) in which the cleaning liquid is stored.
Pierce each two.

[2] Next, the clamps 24 and 44 are respectively opened to open the internal flow paths of the tubes 21 and 41, and the whole blood in the blood bag is captured through the tube 21 and the platelets are captured. Whole blood after being supplied into the filter 7 and passing through the platelet capturing filter 7 (treated liquid)
Is collected into the treated liquid collecting bag 40 via the tube 41.

More specifically, whole blood in the blood bag is transferred through the tube 21 by a drop (self-weight),
Inlet port 8 of the housing 8 (second housing member 82)
21 is supplied into the platelet capture filter 7. When the whole blood passes through the ring 101 of the support member 10b, the flow thereof is uniformly dispersed radially from the central portion of the filter 9, and each porous membrane 921 and the filter member 91 which form the pre-filter member 92. Passing through each of the porous membranes 911 constituting the above. At this time, the platelets are mainly captured by the filter member 91, and other components (unnecessary components) pass through the filter member 91,
It is discharged from the outlet 811 of the housing 8 (first housing member 81) via the support member 10a. And
The processed liquid (whole blood after the platelets have been captured) is transferred through the tube 41 by the drop (self-weight) and collected in the processed liquid collection bag 40. In this way, platelets are filtered and separated from whole blood.

When the platelets have been captured, the clamp 2
Tubes 21, 4 are closed with 4, 44 respectively closed.
The internal channel 1 is closed.

[3] Next, the clamps 34 and 54 are opened, the internal flow paths of the tubes 31 and 51 are opened, and the washing solution in the washing solution bag is passed through the tube 31 and the platelet trapping filter 7 is inserted. The washing liquid (washed liquid) that has been supplied to the inside and passed through the inside of the platelet capture filter 7 is collected into the washed liquid collection bag 50 via the tube 51.

More specifically, the cleaning liquid in the cleaning liquid bag is transferred through the tube 31 by the drop (self-weight),
The platelet trapping filter 7 is supplied from the inflow port 821 of the housing 8 and is supplied to the inside of the housing 8, particularly the filter 9
To wash. As a result, unnecessary components (especially red blood cells and white blood cells) adhering to the inner surface of the housing 8 and the filter 9 are washed away and discharged from the outlet 811 of the housing 8. Then, the washed liquid is transferred through the tube 51 by the drop (self-weight) and collected in the cleaning liquid collection bag 50.

By carrying out such washing, it is possible to more surely reduce the mixing of unnecessary components into the obtained platelet factor. Note that such washing is optional and can be omitted.

When the inside of the platelet capture filter 7 has been washed, the clamps 34 and 54 are closed,
The internal flow paths of the tubes 31, 51 are closed.

[4] Next, the first three-way stopcock 62 and the second three-way stopcock 64 are operated so that the internal space of the syringe 61 and the internal space of the syringe 63 communicate with each other.

In this embodiment, it is assumed that the syringe 61 contains a liquid for stimulating platelets.

In this state, when the plunger of the syringe 61 is moved in the direction in which the volume of the internal space decreases,
The liquid is discharged from the syringe 61, supplied to the platelet capture filter 7 through the first three-way stopcock 62. The liquid comes into contact with the platelets captured by the filter 9 in the platelet capture filter 7. The flow rate of the liquid is set in the range as described above.
This activates the platelets and releases the platelet factor.

The platelet factor released from the platelets is discharged from the outlet 811 of the housing 8 together with the liquid, and is stored in the syringe 63 via the second three-way stopcock 64. At this time, the volume of the internal space of the syringe 63 increases, and the plunger of the syringe 63 also moves accordingly.

Such an operation (movement of liquid) may be carried out only once, but for example, it is preferable to carry out at least one reciprocation, more preferably about 3 to 15 reciprocations. As a result, platelets can be more reliably activated, and as a result, the yield of platelet factor can be increased.

Thus, for example, the platelet-derived growth factor (PDG) is added to the syringe 61 or the syringe 63.
F), transforming growth factor (TGF), epidermal growth factor (EGF), insulin-like growth factor (TG)
Collect a liquid containing platelet factors such as F).

The platelet trapping filter and the platelet factor recovery device of the present invention have been described above with reference to the illustrated embodiments, but the present invention is not limited thereto.

Each of the filter member and the pre-filter member is made of, for example, a block-like (lump-like) porous body instead of a stack of a plurality of porous membranes (membrane-like porous bodies). May be

The filter may be composed of only the filter member by omitting the pre-filter member if necessary.

Further, the filter may further include (have) another member different from the filter member and the pre-filter member.

The first container and the second container of the platelet factor recovery means are replaced by syringes,
For example, it may be composed of a flexible bag (container) or the like.

In the above-mentioned embodiment, the blood to be treated has been described by taking whole blood as a representative, but it goes without saying that the present invention can treat, for example, platelet-rich plasma, platelet suspension such as concentrated platelets and the like. Yes.

[0156]

EXAMPLES Next, specific examples of the present invention will be described.

(Examples 1A to 1D) First, a substance (copolymer) for adjusting the zeta potential on the surface of the filter was synthesized.

2-methoxyethyl acrylate (ME
40 g of A) and 12 g of 2- (N, N-dimethylamino) ethyl methacrylate (DMAEMA) were dissolved in 240 g of dimethylformamide (DMF).

To this solution, 52 mg of azoisobutylnitrile (AIBN) was added, and the reaction was carried out at 75 ° C. for 6 hours under a nitrogen atmosphere.

After completion of the reaction, the reaction solution was concentrated, the concentrate was dissolved in about 200 mL of acetone, and the solution was added dropwise to about 3 L of hexane for purification.

After repeating this purification operation three times, the precipitate was dissolved in about 150 mL of methanol, concentrated and vacuum dried to obtain a MEA / DMAEMA copolymer.

In this copolymer, MEA and DMAE
The composition ratio with MA was confirmed to be 75: 2 by NMR.
It was 5.

Next, two kinds of polyester non-woven fabrics (area weight: 30 g / m ² , thickness: 0.25 mm, average pore diameter: 57.
4 μm, and basis weight: 20 g / m ² , thickness: 0.15 m
m, average pore diameter: 10.7 μm), and the polyester nonwoven fabric was dipped in a methanol solution of 1% of the copolymer. The average pore size of the polyester non-woven fabric was measured with a palm porometer (manufactured by PMI).

After the completion of the immersion, shower washing was performed with RO water kept at 50 ° C. and drying at 70 ° C. to obtain a polyester nonwoven fabric coated with the above copolymer (coated polyester nonwoven fabric).

The zeta potential of the surface of the coated polyester non-woven fabric was measured by using a 0.001N KCl aqueous solution as a medium liquid and a streaming potential measuring device (“ZP2” manufactured by Shimadzu Corporation).
It was 34.1 mV when measured at 0H ").

Each of the coated polyester nonwoven fabrics was cut into a diameter of 19 mm.

In the housing, 7 sheets of coated polyester non-woven fabric having an average pore diameter of 57.4 μm were used as a pre-filter member on the inlet side, and 56 sheets of coated polyester non-woven fabric having an average pore diameter of 10.7 μm were used as a filter member on the outlet side. (Effective membrane area: 2.27 cm ² , volume: 2.27 cm ³ , volume ratio of the filter member to the entire filter: 82.8).
%).

On both sides of the filter, silicone rubber rings (outer diameter: 17 mm, inner diameter: 15 mm, thickness:
0.5 mm) and a polypropylene mesh (thickness: 0.25 mm) inserted in the ring,
A platelet-capturing filter as shown in was prepared.

Furthermore, four platelet factor recovery devices as shown in FIG. 1 were assembled using this platelet capture filter.

Example 2 A coated polyester nonwoven fabric was obtained in the same manner as in Example 1 except that the polyester nonwoven fabric was coated with a 0.5% methanol solution of the copolymer. A capture filter was created and a platelet factor recovery device was assembled.

The coated polyester nonwoven fabric had a surface zeta potential of 29.6 mV.

(Example 3) In the same manner as in Example 1,
An MEA / DMAEMA copolymer was obtained.

In this copolymer, MEA and D
The composition ratio with MAEMA was confirmed by NMR to be 7
It was 5:25.

A coated polyester non-woven fabric was obtained in the same manner as in Example 1 except that the polyester non-woven fabric was coated with a 0.5% methanol solution of the copolymer to prepare a platelet trapping filter. , The platelet factor recovery device was assembled.

The coated polyester nonwoven fabric had a surface zeta potential of 27.4 mV.

(Example 4) In the same manner as in Example 1,
An MEA / DMAEMA copolymer was obtained.

In this copolymer, MEA and D
The composition ratio with MAEMA was confirmed by NMR to be 7
It was 5:25.

A coated polyester non-woven fabric was obtained in the same manner as in Example 1 except that the polyester non-woven fabric was coated with a 1.5% methanol solution of the copolymer to prepare a platelet trapping filter. , The platelet factor recovery device was assembled.

The coated polyester nonwoven fabric had a surface zeta potential of 37.2 mV.

(Embodiment 5) In the same manner as in Embodiment 1,
An MEA / DMAEMA copolymer was obtained.

In this copolymer, MEA and D
The composition ratio with MAEMA was confirmed by NMR to be 7
It was 5:25.

Next, a polyurethane foam (unit weight: 130
g / m ² , thickness: 0.6 mm, average pore size: 8.0 μm) were prepared, and a part of the polyurethane foam was immersed in a methanol solution containing 0.2% of the copolymer. In addition,
The average pore size of the polyurethane foam was measured in the same manner as in Example 1 above.

After completion of the immersion, shower washing was performed with RO water kept at 50 ° C. and drying at 70 ° C. to obtain a polyurethane foam coated with the above copolymer (coated polyurethane foam).

The surface zeta potentials of the coated polyurethane foam and the uncoated polyurethane foam were measured in the same manner as in Example 1 above, and were 33.5 mV and 4.7 mV, respectively.

These coated polyurethane foam and uncoated polyurethane foam were each cut into a diameter of 19 mm.

In the housing, two uncoated polyurethane foams were installed on the inlet side as pre-filter members, and eight coated polyurethane foams were installed on the outlet side as filter members (effective membrane area). : 2.27 cm ² , volume: 1.36 cm ³ , volume ratio of the filter member to the entire filter: 80.0
%).

In addition, on both sides of the filter, as in Example 1 above.
A silicone rubber ring similar to the above and a polyamide mesh were attached to prepare a platelet trapping filter as shown in FIG.

Furthermore, a platelet factor recovery device as shown in FIG. 1 was assembled using this platelet capture filter.

(Comparative Example 1) In the same manner as in Example 1,
An MEA / DMAEMA copolymer was obtained.

In this copolymer, MEA and D
The composition ratio with MAEMA was confirmed by NMR to be 7
It was 5:25.

A coated polyester non-woven fabric was obtained in the same manner as in Example 1 except that the polyester non-woven fabric was coated with a 0.3% methanol solution of such a copolymer to prepare a platelet trapping filter. , The platelet factor recovery device was assembled.

The coated polyester nonwoven fabric had a surface zeta potential of 24.9 mV.

(Comparative Example 2) In the same manner as in Example 1,
An MEA / DMAEMA copolymer was obtained.

In this copolymer, MEA and D
The composition ratio with MAEMA was confirmed by NMR to be 7
It was 5:25.

A coated polyester non-woven fabric was obtained in the same manner as in Example 1 except that the polyester non-woven fabric was coated with a 2.0% methanol solution of such a copolymer to prepare a platelet trapping filter. , The platelet factor recovery device was assembled.

The coated polyester nonwoven fabric had a surface zeta potential of 39.2 mV.

(Comparative Examples 3A to 3D) A filter used in a commercially available leukocyte-removing filter for red blood cell preparation (“Sepacel RZ” manufactured by Asahi Medical Co., Ltd.) was prepared (Basis weight: 41.3 g / m ² , thickness). : 0.2 mm, average pore diameter: 3.
0 μm). The average pore size of the filter was measured in the same manner as in Example 1 above.

The zeta potential on the surface of the filter was measured in the same manner as in Example 1 and found to be 45.0 mV. This filter was cut to a diameter of 19 mm.

Two such filters are placed in the housing.
Four pieces were installed (effective membrane area: 2.27 cm ² , volume:
1.09cm ^3).

In addition, on both sides of the filter, the composition of Example 1
The same silicone rubber ring and polypropylene mesh were attached to prepare a platelet capture filter as shown in FIG.

Further, four platelet factor recovery devices as shown in FIG. 1 were assembled using this platelet capture filter.

[Evaluation 1] Using the platelet factor recovery devices of Examples 1A to 4 and Comparative Examples 1 to 3A, the peripheral blood of healthy individuals (donor D1) anticoagulated with citric acid was tested as follows. The PDGF (platelet factor) was recovered by performing the treatment shown in.

First, donor D1 peripheral blood (treatment liquid) 35
After filtering mL with a platelet capture filter, the platelet capture filter was washed with 10 mL of physiological saline.

Then, add 2 mL to the platelet capture filter.
The physiological saline solution was circulated 10 times at a flow rate of 2 mL / sec to obtain 3 mL of a recovered solution.

Examples 1A to 4 and Comparative Examples 1 to 3
The processing time (blood filtration time), the leukocyte passage rate, the platelet capture rate, the PDGF concentration (filtrate, washed solution, collected solution), the total PDGF recovery amount, and the PDGF recovery amount per 10 ⁹ platelets in the platelet factor recovery device of A were measured. Are shown in Table 1, respectively.

As for the leukocyte passage rate and the platelet capture rate, peripheral blood (treatment liquid) and filtrate (treatment liquid) were sampled, and the white blood cell count and platelet count contained in them were analyzed by multi-item automatic blood cell analysis. Device (Sysmex
"Sysmex SE-9000" manufactured by the same company) was used for the measurement, and the calculation was performed based on the measurement results.

The PDGF concentration was measured by the ELISA method (R
& D Systems, "Quantikine
human PDGF kit ”.

[0208]

[Table 1]

From the results shown in Table 1, all of the platelet factor recovery devices of the present invention (Examples 1A to 4) in which the zeta potential on the surface of the filter member is set to 26 to 38 mV have a short treatment time. Met.

Further, in the platelet factor recovery device of the present invention,
In both cases, the platelet capture rate is extremely high and the leukocyte passage rate is extremely high (leukocyte capture rate is extremely low), and as a result, the platelets are prevented from being activated due to clogging of the filter member with leukocytes. It was confirmed that the platelet factor could be suppressed and that the contamination of the platelet factor into the filtrate and the washed liquid was extremely low.

Further, in the platelet factor recovery device of the present invention,
It was confirmed that platelet factors can be efficiently recovered by physical stimulation.

In particular, the present invention (Examples 1A and 2) in which the zeta potential on the surface of the filter member is set to 29 to 35 mV.
In all of the platelet factor recovery devices, the above results were more excellent.

On the other hand, in the platelet factor recovery device of Comparative Example 1, the platelet capture rate was extremely low due to the low zeta potential on the surface of the filter member.

On the other hand, in the platelet factor recovery devices of Comparative Examples 2 and 3A, the zeta potential on the surface of the filter member was high, so that the leukocyte passage rate was extremely low, that is, the leukocyte trapping on the platelet trapping filter was excessive. It happened.

From the above, it was confirmed that the platelet factor recovery devices of Comparative Examples 1 to 3A could not efficiently recover the platelet factor.

[Evaluation 2] Example 1B and Comparative Example 3B,
Using the platelet factor recovery device of 3C, respectively, to healthy human (donor D2) peripheral blood anticoagulated with citric acid,
The following treatment was performed to collect PDGF (platelet factor).

(Example 1B) First, 35 mL of donor D2 peripheral blood (solution to be treated) was filtered with a platelet capture filter, and then the platelet capture filter was washed with 5 mL of physiological saline.

Then, add 2 mL to the platelet capture filter.
The physiological saline solution was circulated 10 times at a flow rate of 2 mL / sec to obtain 3 mL of a recovered solution.

(Comparative Example 3B) First, 35 mL of donor D2 peripheral blood (treatment liquid) was filtered with a platelet capture filter, and then the platelet capture filter was washed with 10 mL of physiological saline.

Next, a thrombin-added HEPES aqueous solution prepared to give thrombin 1U / 10e ⁸ platelets was added to the platelet-capturing filter and allowed to stand for 10 minutes, after which the HEPES buffer solution was recovered and 3 mL of the recovery solution was collected.
Got

(Comparative Example 3C) First, 35 mL of donor D2 peripheral blood (treatment liquid) was filtered with a platelet capture filter, and then the platelet capture filter was washed with 10 mL of physiological saline.

Next, thrombin 1 U / mL thrombin-added HEPES buffer 24 was added to the platelet capture filter.
25 mL of the collected liquid was obtained by collecting the liquid while passing mL.

Treatment time (blood filtration time) in the platelet factor recovery apparatus of Examples 1B and Comparative Examples 3B and 3C, leukocyte passage rate, platelet capture rate, PDGF concentration (filtrate, washed solution, recovered solution), total PDGF recovery amount. And platelets 10 ⁹
The amount of PDGF recovered per piece is shown in Table 2.

The white blood cell passage rate, platelet capture rate and PDGF concentration were measured in the same manner as in Evaluation 1 above.

[0225]

[Table 2]

From the results shown in Table 2, the present invention (Example 1)
With the platelet factor recovery device of B), it was possible to obtain a recovery amount of platelet factors greater than that in the case of thrombin stimulation (Comparative Examples 3B and 3C) by recovery of platelet factors by physical stimulation.

[Evaluation 3] Using the platelet factor recovery devices of Example 1C and Comparative Example 3D, the peripheral blood of healthy human (donor D3) anticoagulated with citric acid was subjected to the following treatments. PDGF (platelet factor) was recovered.

Example 1C First, 50 mL of donor D3 peripheral blood (solution to be treated) was filtered with a platelet capture filter, and then the platelet capture filter was washed with 10 mL of physiological saline.

Then, add 2 mL to the platelet capture filter.
The physiological saline solution was circulated 10 times at a flow rate of 2 mL / sec to obtain 3 mL of a recovered solution.

(Comparative Example 3D) First, 50 mL of donor D3 peripheral blood (treatment liquid) was filtered with a platelet capture filter, and then the platelet capture filter was washed with 10 mL of physiological saline.

Next, a thrombin-added HEPES buffer solution prepared to give thrombin 1U / 10e ⁸ platelets was added to the platelet-capturing filter and allowed to stand for 10 minutes, and then the HEPES solution was recovered to obtain 3 mL of the recovered solution. It was

Treatment time (blood filtration time), leukocyte passage rate, platelet capture rate, PDGF concentration (filtrate, washed solution, etc.) in the platelet factor recovery apparatus of Example 1C and Comparative Example 3D.
The recovered liquid), the total amount of PDGF recovered, and the amount of PDGF recovered per 10 ⁹ platelets are shown in Table 3, respectively.

The white blood cell passage rate, platelet capture rate and PDGF concentration were measured in the same manner as in Evaluation 1 above.

[0234]

[Table 3]

From the results shown in Table 3, the present invention (Example 1)
In the platelet factor recovery device of C), even when the volume of liquid to be treated (peripheral blood volume) is 50 mL, it is 35 mL (Evaluation 1).
The same performance was confirmed.

On the other hand, in the platelet factor recovery apparatus of Comparative Example 3D, the performance is obviously higher when the amount of liquid to be treated is 50 mL than when it is 35 mL (see Comparative Example 3B in Evaluation 2). It was something that declined.

[Evaluation 4] Using the platelet factor recovery devices of Examples 1D, 5 and 6, the following treatment was carried out on the peripheral blood of healthy individuals (donor D4) anticoagulated with citric acid. PDGF (platelet factor) was recovered.

First, donor D4 peripheral blood (treatment liquid) 10
After filtering mL with a platelet capture filter, the platelet capture filter was washed with 10 mL of physiological saline.

Then, add 2 mL to the platelet capture filter.
The physiological saline solution was circulated 10 times at a flow rate of 2 mL / sec to obtain 2 mL of a recovered solution.

Treatment time (blood filtration time) in the platelet factor recovery apparatus of Examples 1D, 5 and 6, leukocyte passage rate, platelet capture rate, PDGF concentration (filtrate, washed solution, recovered solution), total amount of PDGF recovered and platelets. Table 4 shows the amount of PDGF recovered per 10 ⁹ cells.

The white blood cell passage rate, platelet capture rate and PDGF concentration were measured in the same manner as in Evaluation 1 above.

[0242]

[Table 4]

From the results shown in Table 4, all of the platelet factor recovery devices of the present invention (Examples 1D, 5 and 6) in which the zeta potential on the surface of the filter member was set to 26 to 38 mV had a short treatment time. Met.

Further, in the platelet factor recovery device of the present invention,
In both cases, the platelet capture rate is extremely high and the leukocyte passage rate is extremely high (leukocyte capture rate is extremely low), and as a result, the platelets are prevented from being activated due to clogging of the filter member with leukocytes. It was confirmed that the platelet factor could be suppressed and that the contamination of the platelet factor into the filtrate and the washed liquid was extremely low.

In the platelet factor recovery device of the present invention,
It was confirmed that platelet factors can be efficiently recovered by physical stimulation.

<Summary> The platelet factor recovery device (platelet capture filter) of the present invention was capable of capturing platelets in a short time.

Further, in the present invention, since the concentration of the platelet factor in the filtrate and the washed liquid is extremely low, it is possible to suppress the stimulation of platelets due to the clogging of the filter during filtration. Was confirmed.

Further, in the present invention, since platelets can be captured while suppressing stimulation to platelets, platelets can be easily activated only by physical stimulation,
It was possible to reliably recover a sufficient amount of platelet factor.

On the other hand, in the platelet factor recovery device (platelet trapping filter) of the comparative example, the platelet trapping rate was extremely low, or the platelet trapping rate was sufficient, but the leukocyte trapping rate was extremely high. .

Especially in the latter case, it was confirmed that the blood filtration time becomes extremely long and the platelets are stimulated by the clogging of the filter with the passage of time, which causes the platelet factor to flow out into the filtrate or the washed liquid. It was Further, in this case, since the platelets captured by the filter were already stimulated, it was not possible to recover a sufficient amount of platelet factor even by the thrombin stimulation.

[0251]

As described above, according to the present invention, platelets can be captured more selectively, and a sufficient amount of platelet factor can be recovered from platelets easily and in a short time.

The above effect can be further improved by setting the zeta potential or average pore diameter of the surface of the filter member, installing the pre-filter member, installing the third line for supplying the cleaning liquid, and the like.

Further, in the present invention, aseptic processing, which is an advantage of the filtration method, is possible, and a large amount and rapid processing are possible.

[Brief description of drawings]

FIG. 1 is a schematic diagram showing the overall configuration of a platelet factor recovery device of the present invention.

FIG. 2 is an exploded perspective view (partially shown in section) showing the configuration of the platelet capture filter of the present invention.

[Explanation of symbols]

1 Platelet factor recovery device 2 First line 21 tubes 22 needle tubes 24 clamps 3 third line 31 tubes 32 needle tubes 33 branch connector 34 Clamp 4 second line 40 Processed liquid collection bag 41 tubes 44 clamp 5 fourth line 50 Washed liquid collection bag 51 tubes 53 branch connector 54 Clamp 6 Platelet factor recovery means 61 syringe 62 First three-way stopcock 63 syringes 64 Second three-way stopcock 7 Platelet capture filter 8 housing 81 First Housing Member 811 Outlet 812 lumen 813 Reduced diameter part 814 Mating part 82 Second housing member 821 Inlet 822 lumen 823 Reduced diameter part 824 Mating part 9 filters 91 Filter member 911 porous membrane 92 Pre-filter member 921 porous membrane 10a, 10b Support member 101 ring 102 mesh

   ─────────────────────────────────────────────────── ─── Continued front page    (72) Inventor Tadashi Samejima             1500 Inoguchi, Nakai-cho, Ashigarakami-gun, Kanagawa Prefecture             Terumo Corporation F-term (reference) 4C077 AA13 BB02 CC06 EE01 KK13                       KK25 LL02 LL13 LL15 LL23                       NN02 PP07 PP08 PP09 PP10                       PP12 PP13 PP14 PP16

Claims (11)

[Claims] 1. A platelet trapping filter for trapping platelets from a liquid to be treated containing platelets, which is made of a porous material in a housing having an inlet and an outlet, and has a surface zeta potential of 26. A platelet trapping filter comprising a filter containing a filter member of ~ 38 mV. 2. The surface of the filter member is coated with a copolymer mainly containing an alkoxyalkyl (meth) acrylate and an alkylaminoalkyl (meth) acrylate, whereby the surface has a zeta potential of 26 to 40.
The platelet capture filter according to claim 1, which has a voltage of 38 mV. 3. A platelet trapping filter for trapping platelets from a liquid to be treated containing platelets, which is formed of a porous body in a housing having an inlet and an outlet, and the surface of which is alkoxyalkyl (meth ) A platelet trapping filter comprising a filter containing a filter member coated with a copolymer mainly containing acrylate and alkylaminoalkyl (meth) acrylate. 4. The platelet capture filter according to claim 1, wherein the filter member has an average pore diameter of 5 to 20 μm. 5. The platelet capture filter according to claim 1, a first line connected to the inflow port, for supplying the liquid to be treated into the platelet capture filter, A second line connected to the outlet for collecting the liquid that has passed through the platelet trapping filter, and a platelet factor for activating the platelets trapped by the platelet trapping filter and collecting the platelet factor released from the platelets. A platelet factor recovery device comprising: a recovery means. 6. The platelet factor recovery device according to claim 5, wherein the platelet recovery means activates the platelets by physical stimulation. 7. The platelet factor recovery device according to claim 5, wherein the platelet factor recovery means is configured to share a part of the first line and / or the second line. 8. The platelet factor recovery means has a pair of containers installed via the platelet capture filter,
The platelet factor recovery device according to any one of claims 5 to 7, wherein the first container is connected to the first line and the second container is connected to the second line. 9. A first flow path switching device is installed in the middle of the first line, and the first container is connected to the first line via the first flow path switching device. The platelet factor recovery device according to claim 8, which is connected. 10. A second flow path switching device is installed in the middle of the second line, and the second container is connected to the second line via the second flow path switching device. The platelet factor recovery device according to claim 8 or 9, which is connected. 11. A liquid for stimulating platelets is moved between the first container and the second container to bring them into contact with the platelets captured by the platelet capture filter, The platelet factor recovery device according to claim 8, which activates the platelets.