Cryoprotective agent of peripheral blood mononuclear cells and preservation method of cryoprotective agent

Abstract

The invention discloses a cryoprotective agent of peripheral blood mononuclear cells. The cryoprotective agent is prepared from the following ingredients in parts by weight: 43-53 parts of a culture medium, 30-40 parts of auto-plasma, 12-17 parts of dimethyl sulfoxide, 5-7% of human serum albumin, and the balance being 0.17-0.23mol / L trehalose. The invention discloses a preservation method of the cryoprotective agent. The preservation method comprises the steps of collecting fresh anticoagulation blood, preparing peripheral mononuclear cells into cell suspension by using human serum albumin, adding the cryoprotective agent with the same volume as that of the cell suspension, mixing uniformly, preserving for 10-20h at below 90-below-70-below DEG C, and preserving in liquid nitrogen. According to the cryoprotective agent and the preservation method thereof, a cryopreserved cell has the high motility rate and the strong cell activity and can shorten the Cytokine Induced Killer (CIT) cell induced amplification period.

Description

technical field [0001] The invention relates to the technical field of biological cell preservation, in particular to a cryoprotectant and preservation method for human peripheral blood mononuclear cells. Background technique [0002] Human peripheral blood mononuclear cells refer to cells with a single nucleus in peripheral blood, including lymphocytes, monocytes, dendritic cells and other cells. Human peripheral blood mononuclear cells are the seed cells for the preparation of cytokine-induced CIK cells, and also the seed cells for inducing specific lymphocytes. In actual clinical use, human peripheral blood mononuclear cells are usually extracted from healthy patients for cryopreservation, and then induced and expanded when needed. Therefore, the quality and quantity of human peripheral blood mononuclear cells before and after cryopreservation are directly related To induce the cycle and cell quality of CIK cells, this requires that the damage to the cells be minimized d...

AI Technical Summary

Problems solved by technology

[0003] The existing cryopreservation technology of human peripheral blood mononuclear cells mainly uses serum, osmotic cryoprotectant and culture medium to prepare a cryopreservation solution to freeze the cells; or add hydroxyethyl starch to the cryopreservation solution. Methods The cryopreserved cells may cause allergic reactions during the reinfusion process, and after resuscitation, the cells have a high survival rate but low cell activity, mainly manifested by more irregular cells, and the subsequent induction of culture killing In cell process, the amplification cycle is longer

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