Non-programmable-controlled cooling cryopreservation method and protection agent for hematopoietic stem cells at minus 80DEG C

Abstract

The invention relates to a non-programmable-controlled cooling cryopreservation method and a protection agent for hematopoietic stem cells at minus 80DEG C. The protection agent consists of 5 parts by volume of reagent A and 2 parts by volume of reagent B, wherein the reagent A is a dimethyl sulfoxide and hydroxyethyl starch solution, wherein the mass percents of the dimethyl sulfoxide and hydroxyethyl starch are 10 to 20 percent and 5.4 to 6 percent respectively, the reagent B is an albumin solution, and the mass percent of the albumin is 20 to 40 percent. The formula of the protection agent is simple, and raw materials are cheap; by adopting the preservation method, the hematopoietic stem cells can be simply and conveniently stored under the minus 80-DEG C non-programmable-controlled cooling condition, the survival rate of the stored cells is higher than 90 percent, and the method is suitable for being clinically popularized and applied and used for preserving the clinic hematopoietic stem cells at the low temperature of minus 80DEG C for a short time.

Description

technical field [0001] The invention belongs to the field of refrigerated storage of stem cells, and in particular relates to a -80°C non-programmed cooling and freezing method and a protective agent for hematopoietic stem cells. Background technique [0002] Hematopoietic stem cells are widely used clinically, and cryopreservation of hematopoietic stem cells is one of the key technologies to ensure the success of hematopoietic stem cell transplantation, so the cryopreservation method is particularly important. At present, clinical cryopreservation of hematopoietic stem cells includes -196°C liquid nitrogen program-controlled cooling preservation and -80°C non-programmable cooling preservation. The former can be preserved for a long time with less cell damage, and can generally be preserved for 23 to 25 years, but the operation is complicated and the equipment is expensive. There is cell agglutination, and the latter can preserve stem cells for 1 to 2 years. The operation is...

AI Technical Summary

Problems solved by technology

At present, the protective agents used for low-temperature storage of stem cells at -80°C include dimethyl sulfoxide (hereinafter referred to as DMSO), CP-1, hydroxyethyl starch, RPMI1640 medium, human serum albumin, etc., and the formulations of different protective agents and The methods of use are quite different, and the preservation effect and storage time of stem cells are also different. The survival rate of most preserved cells is about 80%. It is difficult to guarantee the safety of stem cells. Some cryopreservation methods also have the disadvantage of complicated operation.

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