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Erythrocyte cryopreservation method and cryopreserved erythrocytes

A cryopreservation method and red blood cell technology, applied in the field of red blood cell preservation, can solve the problems of complicated glycerin removal operation, red blood cell cannot be directly reinfused, and low red blood cell survival rate, so as to achieve good microscopic shape, save the process of deglycerolization, and save The effect of adding glycerin

Active Publication Date: 2021-04-09
UNIV OF SCI & TECH OF CHINA
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Even so, red blood cells frozen by low-concentration glycerol rapid cryopreservation still cannot be directly reinfused, and they still face the problem of complicated and difficult glycerol removal before use.
In addition, in the current research, the survival rate of red blood cells after freeze-drying is still relatively low, the technology is not mature enough, and it still faces great difficulties in practice

Method used

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  • Erythrocyte cryopreservation method and cryopreserved erythrocytes
  • Erythrocyte cryopreservation method and cryopreserved erythrocytes
  • Erythrocyte cryopreservation method and cryopreserved erythrocytes

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0033] Embodiment 1: Taking trehalose as cryoprotectant

[0034] 1. Implementation method:

[0035] 1) Solution preparation: the concentration of sodium alginate solution is 1.5% (w / v), the concentration of calcium chloride solution is 0.15M, and the concentration of sodium citrate solution is 0.8M. The cryoprotectant formula is: 1.2M trehalose solution.

[0036] 2) Red blood cell pretreatment: take an appropriate amount of leukocyte-removed red blood cell suspension (stored in a refrigerator at 4°C for less than 15 days), add 0.9% NaCl solution and centrifuge to remove the supernatant, repeat the above steps 3 times to obtain a volume of approximately 80% red blood cell solution. Add an appropriate amount of 0.9% NaCl solution to obtain 25 mL of erythrocyte solution with a packing volume of 40%, and place it in a refrigerator at 4°C for later use.

[0037] 3) Add cryoprotectant: Take 25ml of the above-mentioned pretreated erythrocytes and put them into a cryopreservation t...

Embodiment 2

[0049] Embodiment 2: Take glycerol as cryoprotectant

[0050] 1. Implementation method:

[0051] 1) Solution preparation: the concentration of sodium alginate solution is 1.5% (w / v), the concentration of calcium chloride solution is 0.15M, and the concentration of sodium citrate solution is 0.8M. The formulation of the cryoprotectant is: 10% (w / v) glycerin.

[0052] 2) Red blood cell pretreatment: take an appropriate amount of leukocyte-removed red blood cell suspension (stored in a refrigerator at 4°C for less than 15 days), add 0.9% NaCl solution and centrifuge to remove the supernatant, repeat the above steps 3 times to obtain a volume of approximately 80% red blood cell solution. Add an appropriate amount of 0.9% NaCl solution to obtain 25 mL of erythrocyte solution with a packing volume of 40%, and place it in a refrigerator at 4°C for later use.

[0053] 3) Add cryoprotectant: Take 25ml of the above-mentioned pretreated red blood cells and put them into the cryopreser...

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Abstract

The invention relates to the technical field of erythrocyte preservation, in particular to an erythrocyte cryopreservation method and cryopreserved erythrocytes. An erythrocyte low-temperature protective agent used in the invention is an impermeable protective agent, and under the protection of the impermeable protective agent, erythrocytes are wrapped in a gel shell structure formed by cross-linking of sodium alginate to form fibers containing erythrocytes. Compared with the existing erythrocyte cryopreservation technology, the erythrocyte cryopreservation scheme provided by the invention has the advantages that the tedious glycerol adding and deglyceridation processes are omitted, the survival rate of the frozen human erythrocytes in glycerol-free quick cryopreservation can reach 90% or above, and meanwhile, the erythrocytes can keep a good microscopic form.

Description

technical field [0001] The invention relates to the technical field of red blood cell preservation, in particular to a red blood cell freezing method and frozen red blood cells. Background technique [0002] The cryopreservation of blood is the most widely used and most important part of the preservation of all biological materials. The research on blood preservation is of great significance to military medicine and clinical treatment. In particular, the long-term cryopreservation of human red blood cells is of great importance to human society. Significance, such as natural disaster emergency blood transfusion, rare blood type blood transfusion and research, military reserves, etc., all have great demand for the preservation of human red blood cells. Therefore, it is of great significance to study how to efficiently preserve red blood cells for a long time. [0003] At present, the most commonly used red blood cell preservation methods in clinical practice mainly include: ...

Claims

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Application Information

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IPC IPC(8): A01N1/02C12N5/078
CPCA01N1/021A01N1/0221C12N5/0641
Inventor 赵刚覃显慧
Owner UNIV OF SCI & TECH OF CHINA
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