Pretreating liquid and freeze protecting liquid for cryopreservation of erythrocyte and their application

A technology of cryoprotective fluid and pretreatment fluid, which is applied in the fields of red blood cell pretreatment fluid, red blood cell cryopreservation, and red blood cell cryoprotective fluid.

Inactive Publication Date: 2006-11-08
FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

However, although macromolecular substances can increase the glass transition temperature of the protective solution, they

Method used

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  • Pretreating liquid and freeze protecting liquid for cryopreservation of erythrocyte and their application
  • Pretreating liquid and freeze protecting liquid for cryopreservation of erythrocyte and their application

Examples

Experimental program
Comparison scheme
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Embodiment

[0034] Take 50 ml of venous blood from healthy volunteers, add heparin for anticoagulation, and centrifuge to remove white blood cells and platelets. Mix the washed concentrated erythrocytes with 150 ml of pretreatment solution consisting of basic buffer and small molecule sugar, incubate in a water bath at 4-40°C for 1-4 hours, centrifuge to remove the supernatant, and then transfer to a solution containing 10wt%-40wt After 75 milliliters of the cryoprotectant solution of the macromolecule protective component, store it at -80°C or -196°C for 1 week (or directly add the macromolecule protective component to the pretreated erythrocyte suspension and freeze it). When thawing the cells, the samples were taken out, quickly thawed in a 40°C water bath, and the hemolysis rate of red blood cells and other indicators were measured.

[0035] The pretreatment and cryoprotection process are shown in Table 1.

[0036] Example

Basic buffer

in the pretreatment solut...

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PUM

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Abstract

The present invention discloses cryopreservation method of erythrocyte and its erythrocyte pretreating liquid and erythrocyte freeze protecting liquid. The method includes first hatching erythrocyte in erythrocyte pretreating liquid, then protecting erythrocyte in erythrocyte freeze protecting liquid and finally freezing erythrocyte. The erythrocyte pretreating liquid is basic buffering liquid with small molecular sugar component added, and the erythrocyte freeze protecting liquid is macro molecular protecting liquid with small molecular sugar and albumin component added. The present invention protects the inner erythrocyte membrane and outer erythrocyte membrane to raise erythrocyte recovering rate and lower the hemolysis rate of erythrocyte after thawing.

Description

Technical field: [0001] The invention relates to cell cryopreservation technology, in particular to a method for cryopreservation of red blood cells, and a red blood cell pretreatment solution and a red blood cell cryoprotectant solution used in the method. Background technique: [0002] Blood transfusion therapy is an important treatment method to save the lives of the wounded. Sufficient blood supply is an important guarantee to reduce the casualty rate of the wounded and sick. The clinical method for long-term preservation of red blood cells is cryopreservation at -80°C or -196°C. FDA believes that the cryopreservation period of red blood cells can be as long as 10 years or more. The current deep cryopreservation mainly uses high-concentration glycerol as a protective agent, but complex washing procedures are required to remove glycerol after thawing, which consumes a lot of time and manpower. The current clinical routine glycerin elution time is about 3 hours, which ma...

Claims

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Application Information

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IPC IPC(8): A61K35/18A61K47/26
Inventor 权国波韩颖刘敏霞郭永张亮王艳王捷熙刘安
Owner FIELD OPERATION BLOOD TRANSFUSION INST OF PLA SCI ACAD OF MILITARY
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