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Rapid freezing and thawing process for erythrocyte in refrigerator and freezing protection liquid and scrubbing liquid used by the process

A technology of red blood cells and protective solution, applied in biochemical equipment and methods, application, transportation and packaging, etc., can solve the problems of tedious and time-consuming cryopreservation of reagent red blood cells, achieve convenient storage and application, save test time, and improve The effect of utilization

Inactive Publication Date: 2008-12-31
SHANGHAI BLOOD CENT
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0013] One of the purposes of the present invention is to provide a new simple method for long-term frozen red blood cells, which solves the problems of cumbersome and time-consuming red blood cell cryopreservation technology of existing reagents

Method used

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  • Rapid freezing and thawing process for erythrocyte in refrigerator and freezing protection liquid and scrubbing liquid used by the process
  • Rapid freezing and thawing process for erythrocyte in refrigerator and freezing protection liquid and scrubbing liquid used by the process
  • Rapid freezing and thawing process for erythrocyte in refrigerator and freezing protection liquid and scrubbing liquid used by the process

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0038] Effect of red blood cell cryopreservation method under different DMSO concentrations of embodiment 1 (-20 ℃ refrigerator)

[0039] experimental method:

[0040] 1. Prepare red blood cell freezing solution and washing solution.

[0041] 2. Freezing method: Add 1ml of packed red blood cells and 3ml of cryoprotectant into 10ml test tubes, slowly invert the test tubes several times to mix well, and store in a -20°C refrigerator for 4 hours.

[0042]3. Thawing method: Take out the frozen red blood cells (4ml in total), add 4ml of thawing washing solution, and thaw at room temperature or 37°C. After complete melting, slowly invert the test tube several times to mix well, centrifuge at 4000rpm for 3 minutes, and remove the supernatant. Add another 5ml of thawed washing solution, and slowly invert the test tube several times until the packed red blood cells at the bottom of the test tube are completely suspended. Centrifuge at 4000rpm for 3 minutes, remove the supernatant, a...

Embodiment 2

[0052] The effect of red blood cell cryopreservation method (-80 ℃ refrigerator) under the different DMSO concentrations of embodiment 2

[0053] The experimental method is the same as in Example 1, except that the refrigerator is replaced with a -80°C refrigerator. Red blood cell cryopreservation solution and washing solution formulations are also the same as in Example 1.

[0054] Experimental results: as shown in Table 2

[0055] Table 2 Different DMSO concentrations, stored in a -80°C refrigerator, the proportion of red blood cells lost in each step and the recovery rate of red blood cells

[0056]

[0057] *Thawing time for all experiments was within 20 minutes.

Embodiment 3

[0058] The influence of embodiment 3 different macromolecular substances freezing red blood cell method

[0059] Experimental method is the same as embodiment 1.

[0060] Red blood cell cryopreservation solution formula: (the following percentages are all W / V%)

[0061] Preface

No

DMSO concentration

Dextran

concentration

Hydroxyethyl starch

concentration

pvp

concentration

Lactic acid concentration

Na 2 HPO 4

concentration

1

20％

10％

3％

0.2％

2

20％

5％

3％

0.2％

3

20％

2％

3％

0.2％

4

20％

10％

3％

0.2％

5

20％

5％

3％

0.2％

6

20％

2％

3％

0.2％

[0062] 7

20％

10％

3％

0.2％

8

20％

5％

3％

0.2％

...

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Abstract

The invention relates to a technique for preserving erythrocytes in low temperature refrigerator for a long time. Wherein, erythrocyte frozen protective solution is of high osmotic pressure, and contains macromolecular substances with biocompatibility and DMSO (dimethyl sulfoxide). Washing solution is composed of basically isotonic solution and macromolecular substances with biocompatibility. Frozen stock solution can make the erythrocytes go into the hypertonic condition without severe hemolysis. By using macromolecule colloidal medium, prepared erythrocyte washing solution can make the erythrocytes break away from the hypertonic condition so that the erythrocytes are not in severe hemolytic condition. Then the macromolecular substances are washed away by normal saline, and erythrocyte saline suspension is obtained. When the technique is applied in relevant experiments, the total frozen preservation process lasts about one minute, while the unfreezing and washing processes last about fifteen minutes. Thus, the achievement rate of the erythrocytes is high.

Description

technical field [0001] The invention relates to a technology for long-term preservation of red blood cells. Background technique [0002] In the prior art, red blood cells are first suspended in a high-osmotic cryopreservation solution, wherein the main active ingredient is glycerol or DMSO (dimethyl sulfoxide). [0003] Example: Meryman's solution: 57% glycerol, 3% sodium lactate, 0.2% Na 2 HPO 4 , 0.03% KCl. [0004] Huggins solution: 79% glycerol, 8% glucose, 1% fructose, 0.3% EDTA-2Na. [0005] Krijnen solution: 35% glycerol, 2.9% sorbitol, 0.63% NaCl. [0006] Valeri solution: 35% glycerol, 2.88% mannitol, 0.65% NaCl. [0007] Its working principle is to allow hyperosmotic glycerol to penetrate into the red blood cells to replace the water in them, thereby reducing the formation of ice crystals inside and outside the red blood cells during freezing and protecting the red blood cells from being destroyed. In the process of red blood cell thawing, due to the hyperos...

Claims

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Application Information

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IPC IPC(8): C12N5/08A61J1/05A61M1/02A01N1/02
Inventor 向东范亮峰
Owner SHANGHAI BLOOD CENT
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