JPH06269487A - Method for preparing thawed blood from refrigerated blood - Google Patents

Abstract

PURPOSE:To easily and continuously prepare thawed blood for blood transfusion by taking means which contains hollow fiber membranes and separates a liquid prepd. by mixing a physiological salt soln. with a component contg. blood cells mixed with a hematocyte cell protective agent to the component consisting essentially of the blood cells and the hematocyte cell protective agent extract. CONSTITUTION:The prepn. of the thawed blood from the refrigerated blood is executed by first thawing the component contg. the blood cells mixed with the hematocyte cell protective agent, storing the component in a reservoir 1 and mixing the physiological salt soln. 2 therewith in a mixer 3. The liquid mixture is then separated to the component consisting essentially of the blood cells and the hematocyte cell protective agent extract 7 by the blood cell separator 4 contg. the hollow fiber membranes. In succession, the separated component consisting essentially of the blood cells is introduced into another reservoir 8 and the physiological salt soln. 2 is mixed therewith in the mixer 3. The respective strokes mentioned above are repeated until the hematocyte cell protective agent is no longer detected from the component consisting essentially of the blood cells. Thawed plasma 11 is thereafter added to the component consisting essentially of the blood from which the hematocyte cell protective agent is removed.

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 a thawed blood for blood transfusion by thawing a frozen component-separated blood, removing a blood cell protective agent from a blood cell-containing component and mixing it with plasma. .

[0002]

2. Description of the Related Art Conventionally, as a means for securing blood for transfusion of a patient of a special blood type whose blood is difficult to procure, blood for transfusion is prepared from the patient in advance and self-transfusion using this is performed. There is. On the other hand, in recent years, although diagnostic agents for various pathogenic bacteria in blood for transfusion have been developed, highly safe autotransfusion has been considered as a preventive measure against infections associated with transfusion such as hepatitis and AIDS. Therefore, it is expected that autotransfusion will be more important in future blood transfusions.

Autologous blood transfusion is a method of collecting blood during a period in which a patient has enough physical strength to withdraw blood, storing it until necessary, and transfusing the blood. In consideration of the uncertainty of the blood transfusion time, the blood is preserved by freezing. Is considered to be the most suitable method.

[0004] However, in the conventional blood cryopreservation method, a blood cell protective agent such as glycerin is used to separate the blood cell-containing component and plasma by centrifugation or the like, and to prevent the cell destruction by freezing of the blood cell-containing component. Was added and then stored frozen. In addition, in order to use it for blood transfusion after thawing, it is necessary to repeat the centrifugation operation many times in order to remove the added blood cell protective agent for components including blood cells to below the detection limit amount. Blood loss was not small. Further, the centrifugal separation operation is a batch operation, which is extremely troublesome and cannot be easily performed in a hospital, which is also one of the factors that prevent the spread of autologous blood transfusion.

[0005]

An object of the present invention is to provide a simple system capable of transfusing blood that has been frozen and stored after blood collection in a patient's hospital, and in particular of a cryopreserved blood cell protective agent. Blood cells for transfusion by removing the blood cell protective agent from the added components including blood cells and mixing with plasma,
It is to provide a method of continuously and simply preparing.

[0006]

That is, the method for preparing thawed blood of the present invention comprises (a) a step of thawing a component containing blood cells mixed with a blood cell protective agent, and mixing this with physiological saline. , (B) a step of introducing the obtained mixed solution into a blood cell separation device that has a hollow fiber membrane built-in and separates into blood cell-based components and a blood cell cytoprotective agent extraction solution, and (c) the blood cell separation device A step of introducing a blood cell-mainly separated component into a reservoir, and (d)
A step of mixing physiological saline with a component mainly composed of the separated blood cells and leading to a blood cell separation device; and (e) a step of guiding a component mainly composed of blood cells separated by the blood cell separation device to a reservoir,
(f) a step of mixing the physiological saline and separating the blood cell-based component (d) to (e) until a blood cell cytoprotective agent is no longer detected from the blood cell-based component, and (g) ) A step of adding thawed plasma to the obtained component mainly composed of blood cells from which the blood cell protective agent has been removed.

[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, -80
A component containing blood cells mixed with a blood cell protective agent that has been frozen and stored at a temperature of about ℃ for a long period of time is thawed. A component containing thawed blood cells is stored in, for example, a reservoir 1 and physiological saline 2 is mixed therein. The blood cell protective agent 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. On the other hand, physiological saline is used to extract and remove blood cell protective agents such as glycerin from components containing blood cells. The amount of extracted blood cell protective agent is the ratio of the amount of liquid in the components containing blood cells and the amount of physiological saline added, and the ratio of the amount of liquid in the components mainly composed of blood cells after being separated by the blood cell separator. Determined by Usually, physiological saline is added in an amount of about 0.5 to 10 times the amount of components including blood cells. The physiological saline may be added to the components containing the thawed blood cells stored in the reservoir, or may be mixed by the mixing device 3 on the way from the reservoir to the blood cell separating device. It should be noted that the mixing device referred to here only needs to be capable of uniformly mixing a component containing blood cells and physiological saline, and a device for joining two fluids such as a T-shaped tube or a Y-shaped connector can sufficiently achieve the purpose. It

In the blood cell separation device 4 for separating the mixed solution in which physiological saline is mixed with the components containing blood cells in this manner into a component mainly composed of blood cells and a blood cell cytoprotective agent extract (physiological saline). Lead. It is preferable to continuously supply the mixed solution at a constant flow rate to the blood cell separator by using a pump 5 such as a tubing pump.

The blood cell 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 allows a blood cell cytoprotective agent extract to pass through. Usually, the pore diameter is 0.05 to 2 μm, and 2 l / m ² · hr · mm
A hydrophilic porous hollow fiber membrane having a water permeability of Hg or more is used. The material of the hollow fiber membrane used in the present invention is not particularly limited, and 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 membrane, a membrane that has been previously subjected to a hydrophilic treatment with alcohol or the like or a constant hydrophilic treatment by a known method is used.

The hollow fiber membrane module is used in a system in which the mixed solution is flowed through the hollow portion of the hollow fiber membrane and the blood cell protective agent extract is recovered from the outside of the hollow fiber membrane because the loss of blood cell components is reduced. preferable. However, conversely, it is also possible to use a method in which the mixed solution is flown outside the hollow fiber membrane and the blood cell protective agent extract moves to the hollow portion. Usually, the hollow fiber membrane is filled with physiological saline before the start of blood treatment,
The blood cell separation is started under the transmembrane pressure difference as low as possible so that the blood cell component is not damaged. Specifically, the difference between the pressure P _{1 in} the blood flow path before entering the blood cell separator and the pressure P ₂ in the blood cell protective agent extract storage space of the blood cell separator is set to 500 mmHg or less, and normally the blood cell separator is used. The blood cell protective agent extract is sucked by the pump 6 so that the hematocrit value of the components mainly composed of blood cells released from is about 65. The blood cell protective agent extract 7 separated here is discarded.

On the other hand, the components mainly composed of blood cells concentrated by the blood cell separation device are guided to the reservoir 7. The term "reservoir" as used in the present invention refers to a part having a function of temporarily storing components mainly composed of blood cells released from the blood cell separation device, but does not necessarily require a capacity capable of storing the entire amount, and only one blood cell separation is performed. It may be a flow path of a component mainly composed of blood cells from the device to a blood cell separation device for performing the next treatment.

The physiological saline 1 is mixed again with the blood cell-based component introduced to the reservoir 7, and the blood cell-based component diluted with the physiological saline is again guided to the blood cell separation device.
As the blood cell separator used here, a blood cell separator having the same performance as that used previously can be used. In particular,
As shown in FIG. 1, one blood cell separating apparatus and a plurality of reservoirs 1, 8 and switching valves 9, 10 are prepared, and this method may be carried out by using this blood cell separating apparatus many times. As shown in FIG. 2, a plurality of blood cell separation devices may be connected in series to carry out a component containing blood cells while flowing a continuous flow path.

The operation of mixing such a physiological saline solution and separating the component mainly composed of blood cells and the physiological saline solution in which the blood cell protective agent is extracted by a blood cell separating device is carried out from the component mainly composed of blood cells. Repeat until no protective agent is detected. Depending on the amount of physiological saline added and the amount of liquid in the blood cell-based component, once the blood cell separator is passed, the amount of blood cell protective agent in the blood cell-based component is about 1/5. Diluted to the extent. Therefore, normally, if the step of passing through the blood cell separator is performed about four times, the blood cell protective agent will not be detected.

The thawed blood for transfusion is prepared from the frozen blood by adding and mixing the thawed plasma 12 prepared separately to the component composed mainly of blood cells from which the blood cell protective agent has been removed in this way. It

[0016]

EXAMPLES The present invention will be described more specifically below based on examples. Example An apparatus for preparing thawed blood for transfusion as shown in FIG. 1 was assembled. The blood cell separator includes a polyethylene porous hollow fiber membrane (EHF270T, Mitsubishi Rayon Co., Ltd.) as a separation membrane.
Manufactured by the method of flowing blood into the hollow part of the hollow fiber membrane to collect plasma components from the outside of the hollow fiber membrane (membrane area:
0.3 m ² ) was used.

In the reservoir 1 of this apparatus, 130 ml of a component containing blood cells mixed with thawed glycerin (glycerin concentration 33%) was prepared. This solution was mixed with physiological saline at a volume ratio of 1: 6 and then introduced into the blood cell separator 4 at a rate of 100 ml / min. The transmembrane pressure is 4 in the blood cell separator.
The tubing pump was operated so that the pressure became 0 mmHg, and the physiological saline in which glycerin was extracted was removed at a flow rate of 90 ml / min. The obtained blood cell-based component was led to the reservoir 8. The glycerin concentration in this liquid was about 5%.

After the treatment of the liquid in the reservoir 1 is completed,
After switching the switching valves 9 and 10 and mixing the liquid in the reservoir 8 with physiological saline at a volume ratio of 1: 6, 100 ml / min
To the blood cell separator. Here, the physiological saline in which glycerin was extracted again was removed at a flow rate of 90 ml / min, and the obtained blood cell-based components were introduced into the reservoir 1. The glycerin concentration in this liquid was 0.4%.

When the glycerin removal operation using such a blood cell separator was repeated a total of four times, glycerin was not detected in the obtained blood cell-based components. Thawed blood for transfusion was obtained by mixing separately thawed plasma with a component mainly composed of these blood cells.

[0020]

EFFECTS OF THE INVENTION According to the method of the present invention, it is possible to continuously and conveniently prepare thawed blood for transfusion in which a blood cell protective agent is removed from component-separated blood for freezing and mixed with plasma. , It became technically possible to adopt a system that uses blood stored frozen in a hospital where patients are used for blood transfusion.

Further, since the blood cell separator having the hollow fiber membrane with a small priming volume is used, the blood loss at the time of removing the blood cell protective agent from the component-separated blood can be suppressed to the minimum.

[Brief description of drawings]

FIG. 1 is a flow chart showing a method for preparing thawed blood from frozen blood of the present invention.

FIG. 2 is a flow chart showing another aspect of the method of the present invention.

[Explanation of symbols]

 1, 8 Reservoir 2, 12 Physiological saline solution 3, 13 Mixing device 4, 6, 15 Pump 7, 14 Blood cell protective agent extract 9, 10 Switching valve 11 Plasma

Claims (1)

[Claims] 1. A process of: (a) thawing a component containing blood cells mixed with a blood cell protective agent and mixing physiological saline with it; (b) incorporating the obtained mixed solution into a hollow fiber membrane. Then, a step of leading to a blood cell separation device that separates the blood cell-based component and a blood cell cytoprotective agent extract, and (c) a step of guiding the blood cell-based component that has been separated by the blood cell separation device to a reservoir. , (D)
A step of mixing physiological saline with a component mainly composed of the separated blood cells and leading to a blood cell separation device; and (e) a step of guiding a component mainly composed of blood cells separated by the blood cell separation device to a reservoir,
(f) a step of mixing the physiological saline and separating the blood cell-based component (d) to (e) until a blood cell cytoprotective agent is no longer detected from the blood cell-based component, and (g) ) A method of preparing thawed blood from frozen blood, which comprises a step of adding thawed plasma to the obtained blood cell-free component containing blood cells as a main component.