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Cryoprotectant for red blood cells used immediately after being resuscitated and using method

A cryoprotectant and red blood cell technology, applied in the field of cells, can solve the problems of long recovery time, complicated cryopreservation process, and short storage time, and achieve the effect of short time-consuming, no washing steps, and simple ingredients

Active Publication Date: 2019-01-04
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 cryoprotectant for red blood cells used immediately after being resuscitated and a using method, and relates to the technical field of cells. The cryoprotectant for the redblood cells comprises 0.25-0.3 mol / L of trehalose, 5-7% (v / v) of dimethylsulphoxide and 1-3% (v / v) of human serum albumin. The cryoprotectant for the red blood cells is simple in ingredient, when thecells are cryopreserved, the speed of mixing between the red blood cells and the cryoprotectant for the red blood cells does not need to be controlled strictly, and the red blood cells and the cryoprotectant are mixed directly, the cryopreserving method is simple and convenient, and consumed time is shorted. The red blood cells which are cryopreserved by the cryoprotectant can be used after beingresuscitated and are not required to be washed, troublesome and time-consumed washing steps in a glycerinum cryopreserving method are omitted, and demands of emergency blood users are met.

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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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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