JPH06269497A - Method for preparing component separated blood for refrigeration - Google Patents

Abstract

PURPOSE:To provide the method for continuously and easily preparing collected blood to the component separated blood separated to a component contg. blood corpuscles and plasma for the purpose of preservation under refrigeration. CONSTITUTION:An anti-blood coagulation agent 2 is added to the blood 1 and thereafter, the blood is introduced into a plasma separator 3 contg. hollow fiber membranes. The plasma separated therein is introduced into a plasma bag 6 for preservation under refrigeration. On the other hand, the component contg. the separated blood corpuscles is mixed with liquid contg. a hematocyte cell protective agent in a mixer 7 and is then introduced to a blood bag 7 for preservation under refrigeration, by which the component separated plasma for refrigeration is prepd.

Description

Detailed Description of the Invention

[0001]

BACKGROUND OF THE INVENTION 1. Field of the Invention The present invention relates to a method for preparing component-separated blood in which blood cell-containing components and plasma are separated for cryopreservation of blood.

[0002]

2. Description of the Related Art Conventionally, as means for securing blood for a patient of a special blood type whose blood is difficult to procure, so-called autologous blood transfusion has been performed in which the patient himself prepares blood for transfusion. On the other hand, in recent years, although diagnostic agents for various pathogenic bacteria in blood for transfusion have been developed, they are insufficient as a preventive measure against infections associated with blood transfusion such as hepatitis and AIDS, and have a high degree of safety. Blood transfusion is drawing attention.

[0003] Autologous blood transfusion is a method in which the blood of a patient is collected and stored in advance, and in consideration of the uncertain timing of blood transfusion, storage by freezing is considered to be the most suitable method.

[0004] However, in the conventional blood cryopreservation method, a blood cell-containing component is separated into plasma by a centrifugation method, and the blood cell-containing component is protected by blood cells such as glycerin in order to prevent destruction of blood cells by freezing. After adding the agent, it was stored frozen. In addition, in order to use it for blood transfusion after thawing, it is necessary to repeatedly perform a washing operation with physiological saline to remove added blood cell protective agents for components containing blood cells, specifically, centrifugation operation. Was carried out in. At that time, blood loss was not small, and the centrifugal separation operation was a batch operation, which was extremely troublesome and could not be carried out easily in a hospital, which was also one of the factors that prevented the spread of autotransfusion.

[0005]

SUMMARY OF THE INVENTION It is an object of the present invention to provide a simple system in which a blood storage system for using blood, which has been frozen and stored after blood collection, for transfusion, particularly for self-transfusion, can be implemented in a hospital. Yes, especially the collected blood,
It is an object of the present invention to provide a method for continuously and simply preparing a component-separated blood in which a blood cell-containing component and plasma are separated for cryopreservation.

[0006]

[Means for Solving the Problems] That is, a method for preparing blood for freezing component separation according to the present invention is a method in which an anticoagulant is added to blood, the blood is then incorporated into a hollow fiber membrane, and the blood is converted into blood cells. A step of leading to a plasma separation device that separates the contained component and plasma,
A step of introducing the plasma separated by the plasma separator into a plasma bag; a step of introducing the blood cell-containing components separated by the plasma separator into a mixing device and mixing with a liquid containing a blood cell protective agent; and the mixing device. And a step of introducing a component containing blood cells mixed with the blood cell protective agent into a blood cell bag.

[0007]

The method for preparing frozen component-separated blood according to the present invention will be described below with reference to the flow chart shown in FIG.

In the preparation method of the present invention, first, the anticoagulant 2 is added to the collected blood 1. The anticoagulant may be added by adding a predetermined amount in the blood collection bag, or may be continuously added while the blood collected from the human body is continuously guided to the plasma separation device 3. In this way, the blood to which the anticoagulant has been added is guided to the plasma separation device that separates the blood into components containing blood cells and plasma. It is preferable to continuously supply the plasma separation device at a constant flow rate using a pump 4 such as a tubing pump.

As the anticoagulant, a sodium citrate type is preferable, but conventionally used agents such as heparin can be used without particular limitation.

The plasma separation apparatus used in the present invention has a hollow fiber membrane as a separation membrane and a so-called hollow fiber membrane module can be used. The hollow fiber membrane contained in this hollow fiber membrane module needs to be a porous membrane that does not allow blood cell components to pass through but substantially allows plasma components to pass therethrough, and usually has a pore size of 0.05 to 2 μm and a pore size of 0. 2 l / m ²・ hr ・ m
A hydrophilic porous hollow fiber membrane having water permeability of mHg or more is used. The material of the hollow fiber membrane used in the present invention is not particularly limited, polyolefin such as polyethylene, poly (meth) acrylic acid ester such as polymethylmethacrylate,
Polyamide, cellulose derivative and the like can be used. When the material is a hydrophobic film, a film that has been previously subjected to hydrophilic treatment with alcohol or the like, or subjected to constant hydrophilic treatment by a known method is used.

The hollow fiber membrane module is preferably used in a system in which blood is allowed to flow through the hollow portion of the hollow fiber membrane to recover the plasma component from the outside of the hollow fiber membrane because loss of blood cell components is reduced. However, conversely, it is also possible to use the method in which blood is flowed to the outside of the hollow fiber membrane and plasma moves to the hollow portion. Usually, the hollow fiber membrane is filled with physiological saline before the start of blood treatment, and plasma separation is started under a low transmembrane pressure. The difference between the pressure P _{1 in} the blood flow path before flowing into the plasma separation device and the pressure P _{2 in} the plasma storage space of the plasma separation device is 500 mmHg.
It is good to operate as follows.

In the plasma separation apparatus, the plasma stored in the space outside the hollow fiber membrane is sucked by the pump 5 and introduced into the plasma bag 6 for cryopreservation. In this case, the suction of the pump is performed so that the transmembrane pressure difference is as low as possible so that the blood cell component is not damaged, and the hematocrit value of the blood cell-containing component released from the plasma separation device is usually around 65. So as to suck. A particularly preferable method is a method of controlling the flow rate of plasma introduced into the plasma bag so that the transmembrane pressure difference in the plasma separation device is equal to or lower than a certain value. The plasma introduced into the plasma bag is packed and stored frozen. The plasma equivalent to the extra-membrane priming volume collected in the plasma bag at the initial stage of separation is mostly physiological saline and may be discarded.

On the other hand, the component in which the blood cells are concentrated after passing through the hollow portion of the hollow fiber membrane is guided to the mixing device 7, where it is mixed with the liquid 8 containing the blood cell protecting agent. The blood cell protective agent used in the present invention refers to an agent that functions to protect blood cells from destruction of blood cells due to freezing during freezing, and it is common to use glycerin, but ethylene glycol or the like can also be used. . Generally, 60% glycerin isotonic solution is used, and is added about 1.5 times the volume of blood cells. The mixing device used in the present invention only needs to be capable of uniformly mixing a component containing blood cells and a liquid containing a blood cell protecting agent, and a device for joining two fluids such as a T-shaped tube or a Y-shaped connector is sufficient for the purpose. Is achieved.

In this case, the liquid containing the blood cell protective agent added to the mixing device is controlled by the pump 9 and supplied in proportion to the flow rate of the plasma taken out into the cryopreservation plasma bag,
A liquid containing a blood cell protective agent can be uniformly mixed at a predetermined ratio, which is preferable.

The components containing blood cells mixed with the blood cell protective agent in the mixing device are introduced into the blood cell bag 10 for cryopreservation,
Packed and stored frozen.

The blood thus separated into two components is
Each component is stored frozen until blood transfusion. When this frozen stored blood is used for blood transfusion, after each thaw, for components containing blood cells, the blood cell cytoprotective agent added to it is removed by various methods such as centrifugation, and then thawed plasma. Blood for transfusion is prepared by mixing with the components.

[0017]

EXAMPLES The present invention will be described more specifically below based on examples. Example An apparatus for preparing frozen component-separated blood as shown in FIG. 1 was assembled. The plasma separation device includes a polyethylene porous hollow fiber membrane (EHF270W, Mitsubishi Rayon Co., Ltd.) as a separation membrane.
Manufactured, membrane area 0.1 m ² ) was previously hydrophilized with alcohol,
A system was used in which blood was replaced with physiological saline, blood was flown through the hollow portion of the hollow fiber membrane, and plasma components were recovered from the outside of the hollow fiber membrane.

Using this device, blood 20 in a blood collection bag is
0 ml was introduced into the plasma separator at a rate of 15 ml / min. Heparin solution was added as an anticoagulant. When the tubing pump was operated so that the transmembrane pressure difference of the plasma separator was 40 mmHg, a total of 63 ml of plasma could be collected in the plasma bag at a flow rate of about 5 ml / min. The obtained plasma is red blood cells,
Hemolysis was not observed without blood cells such as platelets.

On the other hand, the concentrated components of blood cells discharged from the plasma separator were mixed with 60% glycerin isotonic solution supplied at an average flow rate of 9.5 ml / min at the T-shaped tube portion, and the blood cells were backed up. Blood cells were collected.

[0020]

EFFECTS OF THE INVENTION According to the method of the present invention, component-separated blood for freezing can be continuously and easily prepared from the collected blood, so that the blood of the patient is frozen and stored in the hospital where the patient is located. This made it possible to technically provide a blood storage system used for autotransfusion. In addition, since the plasma separation device incorporating the hollow fiber membrane can be configured with a small priming volume, it is possible to minimize blood loss when preparing component-separated blood.

[Brief description of drawings]

FIG. 1 is a flow chart showing a method for preparing frozen component-separated blood of the present invention.

[Explanation of symbols]

 1 Blood 2 Anticoagulant 3 Plasma Separation Device 4, 5, 9 Pump 6 Plasma Bag 7 Mixing Device 8 Liquid Containing Hemocytoprotective Agent 10 Blood Cell Bag

Claims (2)

[Claims] 1. A step of introducing an anticoagulant into blood, and thereafter introducing the blood into a plasma separation device which has a hollow fiber membrane built therein and separates blood into components containing blood cells and plasma, and the plasma separation. A step of introducing the plasma separated by the device into a plasma bag; a step of introducing the blood cell-containing components separated by the plasma separation device into a mixing device and mixing with a liquid containing a blood cell protective agent; And a step of introducing a component containing blood cells mixed with a protective agent into a blood cell bag. 2. A step of controlling the amount of plasma introduced into the plasma bag so that the transmembrane pressure difference in the plasma separation device becomes a certain value or less, and a supply flow rate of a liquid containing a blood cell protective agent to the mixing device. The method for preparing frozen component-separated blood according to claim 1, further comprising the step of controlling in proportion to the plasma flow rate into the plasma bag.