A cell cryopreservation medium for dendritic cells

Abstract

The invention discloses a cell cryopreservation medium for dendritic cells. An effective amount of cryoprotective agents, cell stabilizers and anti-setting agents are added into RPMI-1640 culture medium and uniformly mixed to form the cell cryopreservation medium for the dendritic cells. The cryoprotective agent is dimethyl sulfoxide, and the cell stabilizer is human gamma globulin; the anti-setting agent is poly-L-lysine, and the molecular weight range of the poly-L-lysine is 30000 to 70000; and the volume percentage concentration of the dimethyl sulfoxide is 1 to 3 percent. When used for cryopreserving DCs, the cell cryopreservation medium disclosed by the invention can ensure the cryopreservation revival rate and immunocompetence of the DCs, and the cryopreservation revival rate and the immunocompetence are not significantly decreased within one year of cryopreservation; the human gamma globulin as the cell stabilizer can maintain the immunocompetence of the DCs; the poly-L-lysine as the anti-settling agent can prevent the human gamma globulin from settling to the bottom of the cell cryopreservation medium in the process of storage, and moreover, the poly-L-lysine has part of the effect of the cryoprotective agent, and can also help to maintain the immunocompetence of the DCs.

Description

technical field [0001] The invention belongs to the field of cell culture, in particular to a cell cryopreservation solution for dendritic cells. Background technique [0002] A large number of mature dendritic cells (DCs) are often needed clinically, but the natural DCs content in the body is small, which is difficult to meet the needs of clinical applications, and can only be obtained in large quantities through in vitro culture. Mature DCs require a large amount of cytokines to maintain, which is expensive and easy to contaminate, which is not conducive to the clinical application of DCs. Therefore, the prepared DCs can be cryopreserved and revived when needed. [0003] In order to maintain the recovery rate and immune activity of cryopreserved DCs, it is extremely important to choose a suitable cryopreservation method. [0004] At present, the DCs cryopreservation method is two-step cooling using a cryopreservation solution containing 10% dimethyl sulfoxide (DMSO) and 2...

AI Technical Summary

Problems solved by technology

[0005] First, the composition of FCS is complex, which will introduce pollutants and pathogens into DCs, and its safety is not easy to control; there are currently imported cryopreservation solutions that do not contain FCS (such as the Cellbanker cell cryopreservation solution produced by ZENOAQ in Japan), but the price Expensive, the application cost is too high, which limits its clinical use, especially in developing and underdeveloped countries;

[0006] Second, although DMSO is a commonly used cryopreservation agent, it does not mean that it will not cause cryopreservation damage to cells (the cytotoxicity of DMSO is inhibited at deep low temperature, and the action should be quick when resuscitating to wash off the dimethyl methoxide as soon as possible. sulfone, otherwise it will cause severe toxicity to cells; however, those skilled in the art know that no matter how fast the speed is, cytotoxicity is inevitable), this damage limits the further improvement of recovery rate; unless new reagents or Reduce the proportion of DMSO

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