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Serum-free cryopreservation medium and method for peripheral blood monouclear cells

A technology of peripheral blood mononuclear cells, applied in the field of immune cell cryopreservation, peripheral blood mononuclear cells completely serum-free cryopreservation solution, to maintain cell stability, reduce possibility, and ensure the effect of biological activity

Inactive Publication Date: 2018-10-16
上海韵飞生物科技有限公司
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  • Abstract
  • Description
  • Claims
  • Application Information

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Problems solved by technology

[0007] The purpose of the present invention is to provide a completely serum-free cryopreservation solution for peripheral blood mononuclear cells and its method, to solve the problem of the peripheral blood mononuclear cell cryopreservation solution and cryopreservation solution that can maintain high cell activity and realize long-term storage and immediate recovery. method problem

Method used

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  • Serum-free cryopreservation medium and method for peripheral blood monouclear cells
  • Serum-free cryopreservation medium and method for peripheral blood monouclear cells

Examples

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Embodiment 1

[0036] Example 1: Preparation of peripheral blood mononuclear cell cryopreservation solution (recombinant human interleukin-2+polyethylene glycol+trehalose+DMEM / F12 medium+human albumin): by volume content, 0.05g / ml human Serum albumin, 50U / mL recombinant human interleukin-2, 0.1g / mL polyethylene glycol, 0.1g / mL trehalose and 99% DMEM / F12 medium were mixed to obtain peripheral blood mononuclear cells Freeze the solution and store it at 4°C for later use.

Embodiment 2

[0037] Embodiment 2: Preparation of peripheral blood mononuclear cell cryopreservation solution (recombinant human interleukin-2+polyethylene glycol+trehalose+DMEM / F12 medium+human albumin): by volume content, 0.05g / ml human Serum albumin, 100 U / mL recombinant human interleukin-2, 0.2 g / mL polyethylene glycol, 0.15 g / mL trehalose and 98% DMEM / F12 medium were mixed to obtain peripheral blood mononuclear cells Freeze the solution and store it at 4°C for later use.

Embodiment 3

[0038] Example 3: Preparation of peripheral blood mononuclear cell cryopreservation solution (recombinant human interleukin-2+polyethylene glycol+trehalose+DMEM / F12 medium+human albumin): by volume content, 1.0g / ml human Serum albumin, 200U / mL recombinant human interleukin-2, 0.0.3g / mL polyethylene glycol, 0.2g / mL trehalose and 95% DMEM / F12 medium were mixed to obtain peripheral blood mononuclear The cell freezing solution was stored at 4°C for later use.

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Abstract

The invention relates to a serum-free cryopreservation medium and method for peripheral blood monouclear cells. The serum-free cryopreservation medium comprises recombinant human interleukin-2, humanserum albumin, polyethylene glycol, trehalose and a basic culture medium or sodium chloride for injection. The serum-free cryopreservation medium has the advantages that the recombinant human interleukin-2 (IL-2) is added into the cryopreservation medium, so that the activity stability of immune cells cultured by thawed cells can be increased greatly, the high activity of original cells can be kept, the immune cells can well keep the physiological function and biological feature of the thawed cells, and the problem of direct large-scale amplification of the thawed cells can be solved effectively; the polyethylene glycol and the trehalose are added into the cryopreservation medium, the cryopreservation medium can replace an existing dimethyl sulfoxide-based (DMSO-based) cryopreservation medium, the toxicity of the cryopreservation medium to the cells is lowered effectively, cell stability is maintained, the direct application safety of the thawed cells is increased, and the thawed cellscan be directly used.

Description

technical field [0001] The invention relates to the technical field of cryopreservation of cells, in particular to the cryopreservation of immune cells, especially to the cryopreservation of peripheral blood mononuclear cells, in particular to a complete serum-free cryopreservation solution for peripheral blood mononuclear cells and a method thereof. Background technique [0002] As an effective method to preserve cells, cell cryopreservation has been widely used in the field of biology. Cryopreservation of donor peripheral blood mononuclear cells can maintain their viability and function, which is of great significance for both clinical and basic research, especially when used for the study of a single immune cell. Cryopreservation of peripheral blood mononuclear cells can not only solve the difficulty in collecting peripheral blood mononuclear cells during the induction of existing immune cells, but also require multiple blood collections, and can be used for tumor treatme...

Claims

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Application Information

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Patent Type & Authority Applications(China)
IPC IPC(8): A01N1/02
CPCA01N1/0221
Inventor 陈金卢彬彬陶飞龙唐欢何丽婷程征雁
Owner 上海韵飞生物科技有限公司
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