Human peripheral blood immune cell cryopreservation efficient stabilizer, application and cryopreservation method

Abstract

The invention relates to a human peripheral blood immune cell cryopreservation efficient stabilizer, application and a cryopreservation method. The cryopreservation efficient stabilizer comprises thefollowing main components: proline, sucrose, polyvinylpyrrolidone, ethylene glycol, ectoin, polyvinyl alcohol and a basic cell culture solution without phenol red. In the cryoprotectant, the concentration of the proline is 1-2% (W / V), the concentration of the sucrose is 5-6% (W / V), the concentration of the polyvinylpyrrolidone is 3-6% (W / V), the proportion of the ethylene glycol is 5-8% (V / V), theconcentration of the ectoin is 2-5% (W / V), the concentration of polyvinyl alcohol is 1-3% (W / V), and the proportion of the basic cell culture solution without phenol red is 92-95% (V / V). The cryopreservation efficient stabilizer disclosed by the invention does not contain dimethyl sulfoxide (DMSO), does not introduce exogenous animal-derived protein, reduces the possibility of animal-derived pollution, avoids adverse reactions and toxic and side effects when recovered cells are transfused back to a patient, ensures the safety of clinical application, can be directly used for clinical cellularimmunotherapy, and is safe and effective.

Description

technical field [0001] The invention belongs to the technical field of cell preservation, and relates to a high-efficiency stabilizer for cryopreservation of human peripheral blood immune cells and its application and cryopreservation method. [0002] technical background [0003] Cellular immunotherapy technology is an emerging technology for clinical tumor treatment in recent years, and it is also regarded as one of the most promising technologies. Human peripheral blood immune cells, including lymphocytes, NK cells, monocytes, dendritic cells, etc. And clinical application is increasingly widely valued. In particular, peripheral blood mononuclear cells (PBMC) are also seed cells for the preparation of cytokine-induced killer cells (CIK), LAK cells, natural killer cells (NK), NKT cells, CTL and γδT cells in vitro. The preservation of these cells, especially cryopreservation, is very important for both basic research and clinical application, especially in vitro induced kil...

AI Technical Summary

Problems solved by technology

The disadvantage is that due to the toxic side effects of dimethyl sulfoxide (DMSO), the cells must be washed after recovery, otherwise it is not conducive to the growth of cell culture

The traditional cell cryopreservation method adds fetal bovine serum, which introduces exogenous components and greatly increases the risk of pathogen contamination

According to reports, adding DMSO and animal serum to preserve cells may cause allergic reactions after the cells are reinfused into the human body, which will have a certain impact on human adoptive cell immunotherapy

There are also some scholars who have improved the traditional technology and used β-glucan, hydroxyethyl starch, human serum albumin or autologous plasma mixed with ordinary cell culture medium to freeze autologous peripheral blood immune cells. However, the time period of cell cryopreservation is different, and the survival rate of cells after cryopreservation is still unstable, which cannot meet the needs of clinical treatment.

Foreign countries have developed and launched commercialized serum-free cell cryopreservation solutions, such as CELLBANK, BAMBANKER, LABOBANK and other brands. These products almost do not contain fetal bovine serum and no animal-derived ingredients. The survival rate of cells after cryopreservation and recovery is as high as 80- 90%, but some commercial cryopreservation solutions still contain DMSO components, usually commercialized cell cryopreservation solutions are more expensive on the market

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