JPS62258322A - Method for separating blood component and device therefor - Google Patents

Abstract

PURPOSE:To separate blood components such as erythrocyte, leukocyte, thrombocyte, etc., from whole blood in high efficiency without fear of ATLA infection, by using a specific process. CONSTITUTION:Blood is fractionated to an oligothrombocytic plasma A constituting an upper layer, a liquid mixture B of thrombocyte and leukocyte constituting the intermediate layer and a concentrated erythrocyte liquid C constituting the lower layer, by strong centrifugal treatment. The concentrated erythrocyte liquid C is diluted with a part of the oligothrombocytic plasma A to correct the hematocrit value of he concentrated erythrocyte liquid to <=80% and the mixed liquid B is subjected to weak centrifugal treatment and fractionated into a lower leukocyte layer and an upper concentrated thrombocyte layer. The strong centrifugal treatment is carried out at 1,700-5,500G for 4-10min to separate the solution into an oligothrombocytic plasma layer, a thrombocyte layer, a leukocyte layer and a concentrated erythrocyte liquid layer from the top downward. The weak centrifugal treatment is carried out at 700-600G for 4-10min and the thrombocyte can be easily separated from the leukocyte. EFFECT:The factor VIII can be obtained as a highly concentrated preparation having decreased side effect in high yield.

Description

DETAILED DESCRIPTION OF THE INVENTION 10. Background Technical Field of the Invention The present invention relates to a blood component separation method and an apparatus therefor. To be more specific, red blood cells, white blood cells, and
The present invention relates to a method and apparatus for efficiently separating blood components such as platelets.

PRIOR ART Hemophilia is a genetic disease that causes a bleeding diathesis due to a congenital deficiency of surface fluid coagulation factor V [I+ intrinsic factor or factor II activity in the blood, and is based on a deficiency of factor vII+. Hemophilia A (factor II deficiency, classic hemophilia), and hemophilia B (factor tX deficiency, Chr
iStmaS disease). However, there is currently no radical treatment for hemophilia, and replacement therapy with factor Vl■ for hemophilia A and factor 1X for hemophilia B is performed as needed. .

In other words, in the event of bleeding, start replenishing the deficient coagulation factor as soon as possible, and replenish it in advance if surgery or dental treatment is to be performed. Factor II is unstable and loses its activity upon storage, so fresh blood must be used, whereas factor II is stable and stored blood may be used. However, it is difficult to replenish a sufficient amount of coagulation factors by using whole blood or plasma as is, and for factor For factor Ⅰ, intravenous injection of concentrated preparations such as purified prothrombin complex is widely practiced.

However, among the factor Vl11 preparations, the factor V1 concentrated preparation highly concentrates factor Vl11, so the factor v coagulation activity is deactivated during the manufacturing process, and the recovery rate is approximately 20%.
%, which is a low value. Furthermore, since it is manufactured using the plasma of many people as raw material, there is a high probability of contamination with viruses such as hepatitis. On the other hand, cryoprecipitate is prepared using a simple method from plasma from a small number of people, so the probability of virus contamination is low.
The recovery rate of factor Vll+ coagulation activity also shows a high value. However, since it is a low-concentration preparation, it contains a large amount of fluid and contaminant proteins such as fibrinogen, and when injected in large quantities, it may cause side effects such as an increase in circulating plasma, an increase in fibrinogen concentration, and hemolysis. be.

Conventionally, in order to separate collected blood into red blood cells, white blood cells, platelets, and their various components, a blood component separation device as shown in Figure 10 is used to separate the various components by various operations. is being carried out.

This blood separation device has a blood collection bag (parent bag) 3 equipped with a blood introduction tubular member 2 to which a bottle needle 1 is connected at the tip, a blood component transfer tubular body 4 attached to the tubular body. 4 is connected to the third child bag 5, and from the tubular body 4 via the first branch pipe 6 and the first separation pipe 7 to the first child bag 8, and the second branch pipe 9. It is connected to a second child bag 11 via a second separation tube 10.

When separating blood into its various components using such a blood component separation device, when the entire surface of the blood collection bag 3 collected from a blood vessel, for example a vein, is subjected to gentle centrifugation, a layer of platelet-rich plasma (PRP) is formed. and white blood cells (LC) and m-thickness blood cells (CR).
C) and a mixed layer. After the upper layer of PRP is transferred to the first child bag 8, the blood component transfer tubular body 4 is separated, and the mixed layer remaining in the blood collection bag is used for blood transfusion as it is without separating the LC. P in first child bag 8
By subjecting the PP to strong centrifugation, the upper layer of platelet-poor plasma (PPP) is transferred to the second child bag 11, and the platelet concentrate (PC) remaining in the first child bag 8 is aggregated.
After pelletizing, it is resuspended in plasma to make a platelet preparation. The PPP transferred to the second child bag 11 is frozen, thawed, and centrifuged to obtain cryoprecipitate (AHG), which is further centrifuged to produce a supernatant containing poor lyoplasma (CRP). is transferred to the third child bag 5.

However, in the above method, white blood cells are not particularly separated, so when red blood cells are transfused, 100% white blood cells are also transfused at the same time. Therefore, if these white blood cells (lymphocytes) contained hemophilia antibodies derived from adult hypodermic cells (ATLA), there was a risk of infecting the patient. In particular, this ATLA has a long incubation period, and even if it is carried, there are no symptoms until it develops.
Since I didn't know what to do, I was worried about the infection mentioned above if I donated blood without knowing.

II. OBJECTS OF THE INVENTION Accordingly, an object of the present invention is to provide a novel blood component separation method and apparatus.

Another object of the present invention is to provide a method for separating blood components and an apparatus for the same without fear of ATLA infection. Still another object of the present invention is to provide a blood component separation method and apparatus that can separate blood components into various single components in a closed system.

The purpose of these developments is to perform strong centrifugation on blood to remove the upper layer (A>
Platelet-poor plasma (PPP) and intermediate layer (B) platelets (P
C) and white blood cells (LC), and the lower layer (
C) Fractionate the red blood cell concentrate (PRC) and dilute the red blood cell concentrate shown in (C) with a portion of the platelet-poor plasma shown in (A>) to determine the hematocrit (Ht) value of the red blood cell concentrate. This is achieved by a blood component separation method characterized by correcting to 80% or less, and then subjecting the mixture shown in (B) to gentle centrifugation to separate the white blood cells in the lower layer and the platelet concentrate in the upper layer.

The above-mentioned purpose is also to perform strong centrifugation on blood to remove the upper layer (A).
Platelet-poor plasma (PPP) and intermediate layer (B) platelets (P
C) and white blood cells (LC), and the lower layer (
C) Fractionate the red blood cell concentrate (PRC) and dilute the red blood cell concentrate shown in (C) with a portion of the platelet-poor plasma shown in (A>) to determine the hematocrit (Ht) value of the red blood cell concentrate. Corrected to 80% or less, the mixture shown in (B) was further subjected to gentle centrifugation to separate the lower layer of leukocytes and the upper layer of platelet concentrate, and the remaining platelet-poor plasma was frozen, thawed, and centrifuged. This is achieved by a blood component separation method characterized by fractionating cryoprecipitate and cryoprecipitate-poor plasma by subjecting the blood to cryoprecipitate and cryoprecipitate-poor plasma.

In addition, the present invention is capable of strong centrifugation at 1700G to 5500G.
This is a blood component separation method that is performed for 4 to 10 minutes. Furthermore, the present invention provides a blood component separation method in which weak centrifugation is performed at 70G to 600G for 4 to 10 minutes.

The objective of the invention also includes: a blood collection container connected to a blood collection needle via a blood introduction tubular body; a platelet-poor plasma container connected to the blood collection container via a first blood component distribution means;
a white blood cell container connected to the blood collection container via a second blood component distribution means and connected to the platelet-poor plasma container via a third blood component distribution means; This is achieved by a blood component separation device consisting of a platelet concentrate container connected via No. 4 blood component distribution means.

The objective of the invention also includes: a blood collection container connected to a blood collection needle via a blood introduction tubular body; a platelet-poor plasma container connected to the blood collection container via a first blood component distribution means;
a leukocyte container connected to the blood collection container via a second blood component distribution means and connected to the platelet-poor plasma container via a third blood component distribution means; and a platelet concentrate container connected via a fourth blood component distribution means, and a cryoprecipitate-poor cryoprecipitate plasma container connected to the platelet-poor plasma container via a fourth blood component distribution means. This is achieved using a blood component separation device.

The present invention also provides a blood component separation device comprising a channel closing means with which a fourth blood component distribution means can communicate.

III. Specific Structure of the Invention Next, the present invention will be specifically explained with reference to the drawings. That is, FIG. 1 shows an embodiment of the blood component separation device according to the present invention, in which a blood collection needle 21 is connected to one end of a blood introduction tubular body (blood collection tube) 22, and a cap 36 is attached as necessary. A blood collection container (blood collection bag) 23 is connected to the other end. This blood collection container 23 is connected to a platelet-poor plasma container 29 via a first blood component distribution means formed by a connecting tube 24, a connector 25, and a connecting tube 26.

On the other hand, the blood collection container 23 is connected to a leukocyte container 31 via a second blood component distribution means formed by a connecting tube 24, a connector 25, and a connecting tube 30, and the leukocyte container 31 is , connection tube 30
, connector 25, connecting tube 26, and connecting tube 2
It is connected to a platelet-poor plasma container 29 via a third blood component distribution means formed by 8. Further, the leukocyte container 31 is connected to the platelet concentrate container 3 through a fourth blood component distribution means formed by a connecting tube 32.
Connected to 3.

FIG. 2 shows another embodiment of the present invention, in which a blood component separation device similar to that shown in FIG. The structure is the same except that the third blood component distribution means is formed by the connecting tube 30, the connecting tool 25, and the connecting tube 37.

FIG. 3 shows still another embodiment of the present invention, which is a blood component separation device similar to that shown in FIG. 2, except that the first blood component distribution means is formed by a connecting tube 39. has a similar configuration.

FIG. 4 shows another embodiment of the blood component separation device according to the present invention, in which a tubular body for introducing blood (blood collection tube)
A blood collection needle 21 is connected to one end of the blood collection needle 22, and a cap 36 is fitted thereon if necessary, and a blood collection container (blood collection bag) 23 is connected to the other end. This blood collection container 2
3 is a connecting tube 24, a connecting tool 25, and a connecting tube 2.
It is connected to two platelet-poor plasma containers via a first blood component distribution means formed by a connecting tube 6 and a connecting tube 28. On the other hand, the blood collection container 23 is connected to the white blood cell container 3 through a second blood component distribution means formed by a connecting tube 24, a connecting tool 25, and a connecting tube 30.
1, and the leukocyte container 31 is connected to the platelet-poor plasma container 29 via a third blood component distribution means formed by the connecting tube 30, the connector 25, the connecting tube 26, and the connecting tube 28. It is connected.

The platelet-poor plasma container 29 also includes a connecting tube 28,
It is connected to a container 35 for poor cryoprecipitate plasma via a fifth blood component flow means formed by the connector 27 and the connecting tube 34 . Furthermore, the leukocyte container 31 has a fourth tube formed by the connecting tube 32.
It is connected to a platelet concentrate container 33 via a blood component distribution means.

FIG. 5 shows another embodiment of the present invention, in which a blood component separation device similar to that shown in FIG. The structure is the same except that the third blood component distribution means is formed by the connecting tube 30, the connecting tool 25, and the connecting tube 37.

FIG. 6 shows still another embodiment of the present invention, in which a blood component separation device similar to that shown in FIG. Other than that, the configuration is the same.

FIG. 7 shows another embodiment of the present invention, in which a blood component separation device similar to that shown in FIG. The structure is the same except that the blood component distribution means is formed by a connecting tube 41.

In the blood component separator described above, each container is usually made of a soft thermoplastic resin such as soft vinyl chloride resin or ethylene-vinyl acetate copolymer, but is not limited thereto. Furthermore, although the connecting tubes, connectors, etc. that form each blood component distribution means are usually made of thermoplastic resins such as soft vinyl chloride resin, ethylene-vinyl acetate copolymer, polycarbonate, and polyethylene terephthalate, It is not limited to these.

Further, each container is provided with a mixed injection port that allows communication with the interior by puncturing a bottle needle if necessary, and is covered with a breakable sealing member 42.

Furthermore, each connecting tube is provided with a channel closing means that allows communication, if necessary. As the flow path closing means that allows fluid to flow, any means that normally closes the flow path, but whose closing member opens when necessary to allow fluid to flow can be used. As such a channel closing means, preferably the channel closing means includes a portion to be broken which allows the channel to be opened by breaking. -For example,
For example, as shown in FIG. 8, a hollow tube 47 is formed of a hard synthetic resin, for example, a hard vinyl chloride resin, and has an outer diameter approximately equal to the inner diameter of the liquid port and a tapered outer end. It consists of a solid columnar body 43 having an outer diameter smaller than the inner diameter of the integrally provided liquid passage port and larger than the inner diameter of the tubular body (for example, 24), and is fixed to the inner wall of the liquid passage port 41. . A notch 44 is always provided on the hollow tube 47 side near the boundary between the hollow tube 47 and the columnar body 43. Before use, the solid columnar body 43 prevents communication between the container (eg 23) and the tubular body (eg 24). When in use, the solid columnar body 43 is inserted into the notch 44 with a finger or the like from the outside.
is cut to establish communication between the container and the tubular body through the inside of the hollow tube 47. If necessary, a flat protrusion 45 corresponding to the outer diameter of the solid columnar body is formed on the end face of the solid columnar body to prevent the pieces of the solid columnar body 43 from blocking the communication tube after cutting.

Further, as shown in FIG. 9, the co-infusion port 49 is shielded in the middle by a partition wall 50, but by removing the cover 52 covering the rubber member 51 provided at the tip of the port 49, the bottle needle is inserted into the rubber member 51. By piercing the bottle needle and inserting it into the partition wall 50, the necessary liquid can be circulated, and if the bottle needle is caught, the rubber member 51 prevents backflow. Further, the rubber member 51 may be omitted and the cover 52 may be covered with a breakable sealing member.

①Specific operation of the V0 invention Next, the separation method according to the invention will be specifically explained using the blood component separation device shown in FIG.

The blood (full surface) collected into the blood collection container 23 through the blood collection needle 21 and blood collection tube 22 is subjected to strong centrifugation treatment by passing the container together into a centrifuge.

This strong centrifugation treatment is performed at 1700G to 5500G (G indicates gravitational acceleration) for 4 to 10 minutes, preferably at 2500G.
Performed at ~5500G. This strong centrifugation process separates platelet-poor plasma (PPP) in the upper layer, platelet concentrate (PC) in the first intermediate layer, white blood cells (LC) in the second intermediate layer, and concentrated red blood cells (PRC) in the lower layer. . Although a small amount of platelet-poor plasma remains on top of the first intermediate layer, it essentially consists of a layer of platelets. Further, although a small amount of red blood cells may be present at the bottom of the second intermediate layer, it is substantially made up of a layer of white blood cells. The connecting tube 30 is closed with a clamp (not shown) to avoid contamination of the intermediate layer, and the connecting tube 24 and the connector 25 are removed, leaving a small amount of platelet-poor plasma in the upper layer to secure the plasma in the platelet concentrate. The blood components are separated into a platelet-poor plasma container 29 through the first blood component distribution means formed by the connecting tube 26 and the connecting tube 28, and the connecting tube 26 is stopped with a clamp (not shown), and the connecting tube 3 is separated.
The components of the first and second intermediate layers are removed from the second intermediate layer formed by the connecting tube 24, the connector 25 and the connecting tube 30, together with a small amount of platelet-poor plasma and a small amount of red blood cells. The blood is separated as a mixture into the white blood cell container 31 through the blood component distribution means, and the lower layer concentrated red blood cell remains in the blood collection container 23.

Next, the connecting tube 30 is closed with a clamp, and a portion of the platelet-poor plasma in the platelet-poor plasma container 29 is poured so that the hematocrit value (Ht value) of the red blood cell concentrate is 80% or less, preferably 50 to 80%. By diluting the red blood cell concentrate by returning it to the blood collection container 23 through the first blood component distribution means, plasma is used as a nutritional source for red blood cells, and about 60% or more of white blood cells, of which about 80% or more of lymphocytes are removed. Obtain a red blood cell preparation.

On the other hand, in the connecting tube 26, the blood collection container 23,
It is cut into platelet-poor plasma container 29, and in connection tube 30, blood collection container 23 and white blood cell container 31.
and a platelet concentration container 33.

The leukocyte container 31 and the platelet concentration container 33 are subjected to a mild centrifugation process. This mild centrifugation process is carried out at 70G to 600G.
It is carried out for ~10 minutes. This mild centrifugation process separates the platelet concentrate into the upper layer and the white blood cells into the lower layer, so the upper layer is transferred to the platelet concentrate container 33. Since this leukocyte container 31 has a flow path closing means 47 that can communicate with the leukocyte container 31, there is no possibility that the liquid will inadvertently move to the platelet concentrate container 33 during or after separation. After separating both containers 31 and 33, the platelet concentrate in container 33 is used as a platelet preparation.

Furthermore, the separation method according to the present invention will be specifically explained using the blood component separation device shown in FIG.

Blood (whole blood) collected into a blood collection container 23 through a blood collection needle 21 and a blood collection tube 22 is subjected to strong centrifugation treatment by passing the container together with a centrifugal valve m machine.

This strong centrifugation treatment is performed at 1700G to 5500G (G indicates gravitational acceleration) for 4 to 10 minutes, preferably at 2500G.
Performed at ~5500G. This strong centrifugation process separates platelet-poor plasma (PPP) in the upper layer, platelet concentrate (PC) in the first intermediate layer, white blood cells (LC) in the second intermediate layer, and concentrated red blood cells (PRC) in the lower layer. . Although a small amount of platelet-poor plasma remains on top of the first intermediate layer, it essentially consists of a layer of platelets. Further, although a small amount of red blood cells may be present at the bottom of the second intermediate layer, it is substantially composed of a layer of white blood cells. The connecting tubes 30 and 34 are each closed with clamps (not shown), and the connecting tubes 24 and 34 are connected, leaving a small amount of platelet-poor plasma in the upper layer to avoid contamination of the intermediate layer and to secure the plasma in the platelet concentrate. The blood component is separated into a platelet-poor plasma container 29 through the first blood component distribution means formed by the device 25, the connecting tube 26, the connecting device 27, and the connecting tube 28, and the connecting tube 26 is cleaned with a cleanser (not shown). The components of the second intermediate layer 113 and the second intermediate layer are formed by the connecting tube 24, the connector 25 and the connecting tube 30 together with the small amount of platelet-poor plasma and a small amount of red blood cells. The white blood cell container 31
The red blood cells are separated as a mixture, and the lower layer concentrated red blood cell remains in the blood collection container 23.

Next, the connecting tube 30 is closed with a clamp, and a portion of the platelet-poor plasma in the platelet-poor plasma container 29 is poured so that the hematocrit value (Ht value) of the red blood cell concentrate is 80% or less, preferably 50 to 80%. By diluting the red blood cell concentrate by returning it to the blood collection container 23 through the first blood component distribution means, plasma is used as a nutritional source for red blood cells, and about 60% or more of white blood cells, of which about 80% or more of lymphocytes are removed. Obtain a red blood cell preparation.

On the other hand, in the connecting tube 26, the blood collection container 23,
It is cut into two containers for platelet-poor plasma and a container 35 for cryoprecipitate-poor plasma, and is also separated at the connecting tube 30 into a blood collection container 23, a leukocyte container 31, and a platelet concentration container 33.

Two platelet-poor plasma containers and a cryoprecipitate-poor plasma container 33 are frozen (for example, at -20 to -80°C, preferably -40 to -80°C), then thawed, and
When subjected to centrifugation treatment at G~5500G for 15 to 20 minutes, it will separate into two layers, so the upper layer poor cryoprecipitate plasma is transferred to the cryoprecipitate poor plasma container 35, and the cryoprecipitate is transferred to the platelet poor plasma container 35. leave it in Both containers 2
By separating 9 and 35 at connecting tube 28 or 34, the cryoprecipitate in container 29 is used as an antihemophilic drug drug substance.

The leukocyte container 31 and the platelet concentration container 33 are subjected to a mild centrifugation process. This mild centrifugation process is carried out at 70G to 600G.
It is carried out for ~10 minutes. This mild centrifugation process separates the platelet concentrate into the upper layer and the white blood cells into the lower layer, so the upper layer is transferred to the platelet concentrate container 33. Since this leukocyte container 31 has a flow path closing means 47 that can communicate with the leukocyte container 31, there is no possibility that the liquid will inadvertently move to the platelet concentrate container 33 during or after separation. After separating both containers 31 and 33, the platelet concentrate in container 33 is used as a platelet preparation.

Incidentally, in the above method, the platelet-poor plasma in the platelet-poor plasma container 29 is returned to the blood collection container to serve as a nutritional source for red blood cells, but the amount of platelet-poor plasma decreases accordingly. Therefore, a red blood cell preservation solution is stored in advance in the cryoprecipitate-poor cryoprecipitate plasma container 35, and after being separated into each component after strong centrifugation, this red blood cell preservation solution is transferred to the blood collection container 23. may be mixed. In this case, it is preferable to provide a channel closing means 47 that can also communicate with the cryoprecipitate-poor cryoprecipitate plasma container 35.

Examples of red blood cell preservation solutions include physiological saline containing glucose, adenine, and a colloid osmotic pressure regulator (mannitol).

The above description has been made regarding the blood component separation device shown in FIGS. 1 and 4, but the white liquor component can be separated by performing similar operations with the devices shown in FIGS. 2 to 3 and 5 to 7. Ru.

Next, the present invention will be explained in more detail by giving Examples.

Example 1 A blood component separation device as shown in FIG. 1 was made of soft vinyl chloride resin. In this device, 400 volumes of human blood (whole blood) was collected into a blood collection container 23 with a capacity of 1400 volumes containing 56 volumes of CPD liquid, and then subjected to strong centrifugation at 3,500 G for 5 minutes. The layers were separated in order from the top: a platelet-poor plasma layer, a platelet concentrate layer, a white blood cell layer, and a red blood cell concentrate layer. Next, in order to prevent other components below the second layer from being mixed into the platelet-poor plasma, a small amount of the platelet-poor plasma is left and passed through the first blood component distribution means to a volume of ff1300.
Approximately 200 m to be transferred to the platelet-poor plasma container 29 of RDL.
1), and a portion of the platelet concentrate, white blood cells, and red blood cells in the second and third layers are transferred to the white blood cell container 31 with a capacity of 80 Inl via the second blood component distribution means (about 80 Inl).
0 m1>, this results in approximately 180 m1 in the blood collection container 23.
of red blood cell concentrate remained.

Approximately 30 volumes of platelet-poor plasma in the platelet-poor plasma container 29 was added to this red blood cell concentrate to bring the hematocrit value below 80%. Then, the platelet-poor plasma container 29 was separated from the blood collection container 23. On the other hand, the leukocyte container 31 and the platelet concentrate container 33 are separated from the blood collection container 23, and the
When subjected to mild centrifugation at 0 G for 5 minutes, platelet concentrate was separated in the upper layer and white blood cells were separated in the lower layer, so the upper layer was transferred to the platelet concentrate container 35.

The percentages of white blood cells, red blood cells, and platelets in each component of the blood thus separated were as shown in Table 1.

In the comparative example, the entire surface was centrifuged at 1100G for about 5 minutes using a conventional separation method to separate the red blood cell concentrate and multifaceted platelet plasma, and then the platelet-rich plasma was centrifuged at 3500G.
The results are shown when the platelet concentrate and plasma were separated by centrifugation for about 5 minutes.

In the table, values are shown as inventive/comparative examples.

Table 1 Red blood cells Platelet concentrate Concentrated white blood cells (%) 41/96 <1/
2 Lymphocytes (%) 15/95 <1
/4 Red blood cells (%) 96/100 <
1/<1 platelets (%) 26/34
41/63' Example 2 A blood component separation device as shown in Fig. 4 was made of soft vinyl chloride resin.This device had a CPD of 56d.
After collecting 400w11 of human blood (full surface) into the blood collection container 23 with a capacity of 400m,
When subjected to strong centrifugation at 0 G for 5 minutes, the layers were separated from the top into a platelet-poor plasma layer, a platelet-rich liquid layer, a white blood cell layer, and a red blood cell-rich liquid layer. Next, a small amount of the platelet-poor plasma is passed through the first blood component distribution means to a volume of 30 ml, leaving a small amount of the platelet-poor plasma so that other components below the second layer do not mix into the platelet-poor plasma.
Approximately 200 rI to transfer to the 0 m platelet-poor plasma container 29
IIt), and a portion of the platelet concentrate and white blood cell and red blood cell concentrates in the second and third layers are transferred to the white blood cell container 31 with a capacity of 80 d via the second blood component distribution means (about 80 d).
0m>, and as a result, approximately 180 d of concentrated red blood cell liquid remained in the blood collection container 23.

Approximately 301,112 platelet-poor plasma in the platelet-poor plasma container 29 is added to this red blood cell concentrate to bring the hematocrit value to 80.
% or less. Next, the platelet-poor plasma container 29 and the cryoprecipitate-poor plasma container 35 are transferred to the blood collection container 2.
3, frozen at -60°C and thawed, approximately 25
When centrifuged at 00G for about 20 minutes, poor cryoprecipitate plasma was separated in the upper layer and cryoprecipitate was separated in the lower layer, so the upper layer was transferred to the cryoprecipitate poor plasma container 35 to obtain about 190rn1. About 10rn1 remained in the platelet-poor plasma container 29. On the other hand, when the leukocyte container 31 and the platelet concentrate container 33 were separated from the blood collection container 23 and subjected to mild centrifugation at about 170G for 5 minutes, the platelet concentrate was separated into the upper layer and the white blood cells were separated into the lower layer. was transferred to the platelet concentrate container 33.

The percentages of white blood cells, red blood cells and platelets in each component of the blood thus separated were as shown in Table 2.

In the comparative example, the entire surface was centrifuged at 1100G for about 5 minutes using a conventional separation method to separate the red blood cell concentrate and platelet-rich plasma, and then the platelet-rich plasma was centrifuged at 3500G.
The results are shown when the platelet concentrate and plasma were separated by centrifugation for about 5 minutes.

In the table, values are shown as inventive/comparative examples.

Table 2 Red blood cells Platelet concentrate Concentrated white blood cells (%) 41/96 <1/2
Lymphocytes (%) 15/95 <1/
4 Red blood cells (%) 96/100 <
1/<1 salivary platelet (%) 26/34
41/63 Example 3 Using the same equipment as in Example 2, physiological saline containing glucose, adenine, and a colloid osmotic pressure regulator (mannitol) was added as a red blood cell preservation solution to the cryoprecipitate-poor cryoprecipitate plasma container 33. Example 2
The same procedure as above was carried out, except that after strong centrifugation, 80 m of this physiological saline was transferred to a blood collection container to dilute the red blood cell concentrate.

As a result, the same results as in Example 2 were obtained.

■0Specific Effects of the Invention As described above, the present invention is capable of subjecting blood to strong centrifugation to produce an upper layer (A) of platelet-poor plasma, an intermediate layer (B) of platelets and white blood cells, and a lower layer. (C) a concentrated red blood cell solution, and dilute the concentrated red blood cell fluid shown in (C) with a portion of the platelet-poor plasma shown in (8) to reduce the hematocrit value of the concentrated red blood cell fluid to 80% or less. Correct and further (B)
This patented component separation method is characterized by subjecting the mixed solution shown in the figure to a weak centrifugal process to separate the white blood cells in the lower layer and the platelet concentrate in the upper layer. The platelets are separated into a white blood cell layer and a concentrated red blood cell layer, and due to the cushioning properties of the white blood cell layer, platelets are not subjected to shock and do not aggregate even during strong centrifugation. Furthermore, since platelets can float in plasma contained in the same container, they can be easily separated from leukocytes by mild centrifugation.

This eliminates the need for the agitations required in the conventional method, and provides a platelet preparation whose activity is not impaired. In addition, since there is no need to pellet platelets as in the conventional method, damage to platelets is reduced, and after blood collection, -
A stable preparation can be obtained even after being left for day and night. Therefore, the storage period can be extended. Furthermore, since there are extremely few white blood cells, particularly causative lymphocytes, in the red blood cell concentrate from which white blood cells have been removed by strong centrifugation, there is a possibility that ATLA infection can be prevented.

In addition, the present invention performs strong centrifugation on blood to separate it into (A) platelet-poor plasma in the upper layer, (B) a mixed liquid consisting of platelets and white blood cells in the middle layer, and (C) concentrated red blood cell liquid in the lower layer. death,
The red blood cell concentrate shown in (C) is diluted with a portion of the platelet-poor plasma shown in (△) to correct the hematocrit value of the red blood cell concentrate to 80% or less, and the mixed liquid shown in (B) is further diluted with a portion of the platelet-poor plasma shown in (△). Centrifugation is performed to separate the white blood cells in the lower layer and the platelet concentrate in the upper layer, and the remaining platelet-poor plasma is frozen, thawed, and centrifuged to produce cryoprecipitate and cryoprecipitate-poor plasma. Since this is a blood component separation method characterized by fractionation, in addition to the above-mentioned effects, factor Vll+ can be obtained at an extremely high recovery rate.

Furthermore, since it is obtained as a highly concentrated preparation, it has the advantage of having fewer side effects.

The present invention also provides a blood collection container connected to a blood collection needle via a blood introduction tubular body, a platelet-poor plasma container connected to the blood collection container via a first blood component distribution means, a white blood cell container connected to the blood collection container via a second blood component distribution means and connected to the platelet-poor plasma container via a third blood component distribution means; This is a blood component separation device consisting of a platelet concentrate container connected via the blood component distribution means of No. 4.
Not only can each component of blood be separated in a closed system, but there is no leukocyte contamination, so ATLA
This makes it possible to produce a concentrated red blood cell solution without fear of infection, and also makes it possible to produce a platelet preparation with low platelet damage and stable activity since there is no need for agitations or pelleting.

The present invention also provides a blood collection container connected to a blood collection needle via a blood introduction tubular body, a platelet-poor plasma container connected to the blood collection container via a first blood component distribution means, a white blood cell container connected to the blood collection container via a second blood component distribution means and connected to the platelet-poor plasma container via a third blood component distribution means; A blood component consisting of a platelet 'a condensate liquid container connected via a fifth blood component distribution means, and a platelet concentrate container connected to the platelet-poor plasma container through a fifth blood component distribution means. Since it is a separation device, in addition to the above-mentioned effects, there is an advantage that factor "/II+" can be recovered at an extremely high recovery rate and at a high concentration.

[Brief explanation of drawings]

1 to 7 are front views showing each embodiment of the blood component separation device according to the present invention, FIG. 8 is a cross-sectional view showing an example of a communicating channel closing means, and FIG. 9 is an example of a mixed injection boat. FIG. 10 is a front view showing an example of a conventional blood component separation instrument. 21...Blood collection needle, 22...Tubular body for blood introduction 23...Blood collection container, 24.26, 28.30, 32.34.37°38.3
9.4... Connection tube, 25.27... Connector, 29... Container for platelet-poor plasma, 33... Container for platelet concentrate, 31... Container for white blood cells, 35... Container for scarce lyoprecipitate. rtsu00 ゞ Φ clear~.

Claims (10)

[Claims] (1) Blood is subjected to strong centrifugation and fractionated into (A) platelet-poor plasma in the upper layer, (B) a mixture of platelets and white blood cells in the middle layer, and (C) concentrated red blood cell liquid in the lower layer. The red blood cell concentrate shown in (C) is diluted with a portion of the platelet-poor plasma shown in (A) to bring the hematocrit value of the red blood cell concentrate to 80.
% or less, and the mixture shown in (B) is further subjected to gentle centrifugation to be fractionated into white blood cells in the lower layer and platelet concentrate in the upper layer. (2) Strong centrifugation at 1700G to 5500G for 4 to 10
The method according to claim 1, which is carried out for minutes. (3) The method according to claim 1 or 2, wherein the weak centrifugation treatment is performed at 700G to 600G for 4 to 10 minutes. (4) Blood is subjected to strong centrifugation to separate it into (A) platelet-poor plasma in the upper layer, (B) a mixture of platelets and white blood cells in the middle layer, and (C) concentrated red blood cell liquid in the lower layer. The red blood cell concentrate shown in (C) is diluted with a portion of the platelet-poor plasma shown in (A) to bring the hematocrit value of the red blood cell concentrate to 80.
% or less, and further, the mixture shown in (B) is subjected to gentle centrifugation to separate the white blood cells in the lower layer and the platelet concentrate in the upper layer,
and the remaining platelet-poor plasma is subjected to freezing, thawing, and centrifugation treatment to be fractionated into cryoprecipitate and cryoprecipitate-poor plasma. (5) Strong centrifugation at 1700G to 5500G for 4 to 10
5. The method according to claim 4, wherein the method is carried out for minutes. (6) The method according to claim 4 or 5, wherein the weak centrifugation treatment is performed at 700G to 600G for 4 to 10 minutes. (7) a blood collection container connected to a blood collection needle via a blood introduction tubular body; a platelet-poor plasma container connected to the blood collection container via a first blood component distribution means; a white blood cell container connected to the container via a second blood component distribution means and connected to the platelet-poor plasma container via a third blood component distribution means; and a white blood cell container and a fourth blood component. A blood component separation device consisting of a platelet concentrate container connected via a component distribution means. (8) The blood component separation device according to claim 7, comprising a channel closing means with which the fourth blood component distribution means can communicate. (9) a blood collection container connected to a blood collection needle via a blood introduction tubular body; a platelet-poor plasma container connected to the blood collection container via a first blood component distribution means; a white blood cell container connected to the container via a second blood component distribution means and connected to the platelet-poor plasma container via a third blood component distribution means; and a white blood cell container and a fourth blood component. A blood component separation device comprising a platelet concentrate container connected via a component distribution means, and a cryoprecipitate-poor cryoprecipitate plasma container connected to the platelet-poor plasma container via a fifth blood component distribution means. utensils. (10) The blood component separation device according to claim 9, comprising a channel closing means with which the fourth blood component distribution means can communicate.