Method for separating and culturing peripheral blood mononuclear cells

Abstract

The invention provides a method for separating and culturing peripheral blood mononuclear cells, and relates to the field of biotechnology cell culture. The separation and culture method of the peripheral blood mononuclear cells comprises the following steps: S1, collecting peripheral blood, S2, carrying out centrifugal separation, S3, extracting peripheral blood mononuclear cells, S4, purifying the peripheral blood mononuclear cells, S5, preparing a cell culture medium, S6, carrying out cell culture, and S7, performing preservation. An ALK inhibitor and a GSK-3 inhibitor are arranged, cytopathy in a cell culture process can be inhibited, excessive activation of pathological cells is prevented and rapid proliferation is carried out, animal serum is not added in the culture process, so thatthe possibility of obtaining viruses from animals in the culture process is reduced, the safety in the culture process is ensured, and peripheral blood mononuclear cells are cleaned and purified formultiple times after separation is completed, so that the cultured cells basically do not contain other impurity cells, and the cultured cells are relatively pure.

Description

technical field [0001] The invention relates to the field of biotechnology cell culture, in particular to a method for separating and culturing peripheral blood mononuclear cells. Background technique [0002] Peripheral blood is blood other than bone marrow, which is rich in sources and convenient to collect. Immune cells isolated and cultured from peripheral blood have the characteristics of fast proliferation, safety, persistence, adaptability, and systemicity. Compared with traditional radiotherapy and other methods, active immunization can stimulate systemic anti-tumor effects. More extensive, especially suitable for malignant tumors with multiple lesions or extensive metastases; side effects are minimal. Peripheral blood mononuclear cells are cells in the peripheral blood that have a single nucleus, including lymphocytes and monocytes. [0003] During the culture process of peripheral blood mononuclear cells, due to some changes in light, temperature and environment,...

AI Technical Summary

Problems solved by technology

[0005] Aiming at the deficiencies of the prior art, the present invention provides a method for separating and culturing peripheral blood mononuclear cells, which solves the problem that the cultured cells are prone to disease