High-efficiency NK (natural killer) cell proliferation method

Abstract

The invention relates to the technical field of cell culture, and discloses a high-efficiency NK (natural killer) cell proliferation method. The method is characterized by comprising the following steps: (1) coating a cell culture bottle used for NK cell culture with coating liquid; (2) separating a peripheral blood mononuclear cell (PBMC) containing an NK cell from peripheral blood; (3) adding acell stimulating factor into a cell activation culture solution for PBMC activation; (4) inoculating the activated PBMC into the cell culture bottle after coating treatment in the step (1); and (5) performing induction culture on the PBMC by using an induction culture solution to obtain the NK cell with high activity and purity. The NK cell proliferation method provided by the invention has the characteristics of being simple to operate, high in proliferation efficiency, high in cell activity and high in cell purity; no feeder cells and heterologous serum are added during the culture, therebynot causing medical ethic and GVHD (graft-versus-host disease) risk problems; and the method is applicable to large-scale NK cell preparation and clinical application.

Description

technical field [0001] The invention relates to the technical field of cell culture, in particular to an efficient NK cell expansion method. Background technique [0002] NK cells (Natural killer cells, NK cells) are natural killer cells, an important part of the body's natural immune system, and an important component of anti-virus and anti-tumor cells. [0003] The development of NK cells takes place as early as in the embryonic liver and transfers to the bone marrow after birth. NK cells are a subset of peripheral blood lymphocytes with an immunophenotype, characterized by positive expression of CD56 and lack of CD3 and T cell receptor proteins on the cell surface antigen. Currently, the most common surface markers for detecting NK cells are CD3-, CD16+ and CD56+. [0004] Studies have shown that the number of NK cells in most tumor patients decreases and the ability of NK cells to clear cancer cells decreases. The expression of activating receptors and inhibitory recep...

AI Technical Summary

Problems solved by technology

However, this increases the cell sorting steps, and the currently more effective sorting and separation methods are more expensive, which increases the overall cost of cell culture; secondly, NK cells will also be lost during the sorting process, reducing the number of cells and reducing the cost of cell culture. The large-scale expansion efficiency of cells is not conducive to clinical application

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