A kind of ready-to-use erythrocyte cryoprotectant and its application method

A cryoprotectant and red blood cell technology, applied in the field of cells, can solve the problems of complex freezing process, short storage time, long recovery time, etc., and achieve the effects of eliminating washing steps, short time-consuming and simple components

Active Publication Date: 2021-02-02
SHANDONG QILU STEM CELL ENG
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0005] The invention provides a ready-to-use erythrocyte cryoprotectant for resuscitation and its use method, which overcomes the above-mentioned problems of short storage time, complicated cryopreservation process and long resuscitation time

Method used

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Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0023] (1) Add trehalose, human serum albumin, and DMSO to the aqueous solution in sequence to prepare a red blood cell cryoprotectant, in which the content of DMSO is 6% (v / v), and the concentration of trehalose is 0.28mol / L; human serum albumin The protein content is 2% (v / v);

[0024] (2) Centrifuge fresh blood with added anticoagulant at 4°C and 2500 rpm for 10 minutes, remove the plasma to obtain concentrated red blood cells, mix 10 mL of concentrated red blood cells with 100 mL of red blood cell cryoprotectant, place at room temperature for 30 minutes, and place in gradient In the cooling box, the temperature was gradually lowered to -80°C in a deep-low temperature refrigerator, and the cooling rate was 1°C / min. After 7 days, it was taken out and placed in a water bath at 37°C with constant shaking to recover.

[0025] The recovery rate of erythrocytes determined by hemocytometer counting was 54.69%, and the cell viability of erythrocytes measured by trypan blue detecti...

Embodiment 2

[0027] (1) Add trehalose, human serum albumin, and DMSO to the aqueous solution in sequence to prepare a red blood cell cryoprotectant, in which the content of DMSO is 5% (v / v), and the concentration of trehalose is 0.3mol / L; human serum albumin The protein content is 2% (v / v);

[0028] (2) Centrifuge fresh blood with added anticoagulant at 4°C and 2500 rpm for 10 minutes, remove the plasma to obtain concentrated red blood cells, mix 10 mL of concentrated red blood cells with 100 mL of red blood cell cryoprotectant, place at room temperature for 30 minutes, and place in gradient In the cooling box, the temperature was gradually lowered to -80 °C in a deep-low temperature refrigerator, and the cooling rate was 2 °C / min. After 7 days, it was taken out and placed in a water bath at 37°C with constant shaking to recover.

[0029] The recovery rate of erythrocytes determined by hemocytometer counting was 45.28%, and the cell viability of erythrocytes measured by trypan blue detect...

Embodiment 3

[0031] (1) Add trehalose, human serum albumin, and DMSO to the aqueous solution in sequence to prepare a red blood cell cryoprotectant, in which the content of DMSO is 7% (v / v), and the concentration of trehalose is 0.15mol / L; human serum albumin The protein content is 2% (v / v);

[0032] (2) Centrifuge the fresh blood with added anticoagulant at 4°C and 2500 rpm for 10 minutes, remove the plasma to obtain concentrated red blood cells, mix 10 mL of concentrated red blood cells with 50 mL of red blood cell cryoprotectant, place at room temperature for 30 minutes, and place in gradient In the cooling box, the temperature was gradually lowered to -80°C in a deep-low temperature refrigerator at a cooling rate of 1.5°C / min. After 7 days, it was taken out and placed in a 37°C water bath for recovery with constant shaking.

[0033] The recovery rate of erythrocytes determined by hemocytometer counting was 30.69%, and the cell viability of erythrocytes measured by trypan blue detection...

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Abstract

The invention relates to a ready-to-resuscitate cryoprotectant for erythrocytes and a use method thereof, and relates to the field of cell technology. The composition of the cryoprotectant for erythrocytes provided by the invention includes 0.25-0.3mol / L trehalose, 5-7% (v / v ) of dimethyl sulfoxide, 1~3% (v / v) of human serum albumin. The erythrocyte cryoprotectant provided by the invention has simple components, and it is not necessary to strictly control the mixing speed between the erythrocytes and the erythrocyte cryoprotectant when the cells are frozen, just mix them directly, and the cryopreservation method is simple and time-consuming. The cryopreserved red blood cells of the present invention can be used without washing after resuscitation, which saves the tedious and time-consuming washing steps in the glycerol cryopreservation method and meets the needs of emergency blood users.

Description

technical field [0001] The invention belongs to the field of cell technology, and in particular relates to a ready-to-use red blood cell cryoprotectant and a use method. Background technique [0002] At present, most blood is stored at 2-6°C, and the storage period is about 20 days, which cannot meet the needs of long-term large-scale storage, especially for some rare blood types, such as RH negative blood. Due to the small number of people, Rh-negative blood is often called "panda blood". Therefore, once these patients are ill and urgently need blood transfusion, the Rh-negative blood stock of the blood bank will be put to the test. In addition, some blood donated without compensation is wasted due to the short storage time, but on the other hand, when a major disaster (such as an earthquake, war) requires a large amount of blood, the supply is insufficient. Therefore, the long-term preservation of red blood cells It has important clinical and strategic significance. [0...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 宋振涛陈恒曲廷瑜
Owner SHANDONG QILU STEM CELL ENG
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