Expansion in vitro purification culture method of mesenchymal stem cells and culture medium

Abstract

The invention provides an expansion in vitro purification culture method of mesenchymal stem cells and a cell culture medium used in a culture process. According to the method, mesenchymal stem cells differentiate to osteogenic stem cells. The cell culture medium comprises the following components: 8-12mu g / L of bFGF (Basic Fibroblast Growth Factor), 8-12mu g / L of EGF (Epidermal Growth Factor), 8-12mu g / L of PDGF-BB (Platelet Derived Growth Factor-BB), 10 percent of fetal calf serum, 100U / mL of penicillin and 100mg of L of streptomycin; a solvent is a basic culture medium; and the basic culture medium is a DMEM (Dulbecco Modified Eagle Medium) culture solution, an F12 culture solution or a mixture of the DMEM culture solution and the F12 culture solution. The mesenchymal stem cells obtained by adopting a method comprising the steps of early replacement of the cell culture medium, cell adherence passage and combined addition of cell factors (bFGF, EGF and PDGF-BB) in the culture solution are high in purity and strong in vitro proliferation capacity, and have good capability of differentiation to osteogenic stem cells.

Description

technical field [0001] The present invention claims to protect a method for in vitro expansion and purification of bone marrow mesenchymal stem cells that facilitates the differentiation of bone marrow mesenchymal stem cells into osteoblasts, and the cell culture medium used in the culture process. Background technique [0002] Bone marrow mesenchymal stem cells can be differentiated into osteoblasts, chondrocytes, adipocytes and other cell types under the stimulation of appropriate conditions, and are easy to isolate and culture, and have strong self-renewal, differentiation ability and immunosuppressive advantages. It has important application prospects in the fields of tissue and organ defect diseases, tissue and organ degenerative diseases, cell therapy and tissue engineering. Recently, many domestic and foreign studies have explored the in vitro isolation and culture methods of bone marrow mesenchymal stem cells, mainly including adherent separation, density gradient ce...

AI Technical Summary

Problems solved by technology

Recently, many domestic and foreign studies have explored the in vitro isolation and culture methods of bone marrow mesenchymal stem cells, mainly including adherent separation, density gradient centrifugation and cell sorting. And cell sorting can obtain higher purity cells, but the cell viability is relatively low, and the amount of bone marrow required is large, which affects subsequent research and application

Moreover, the bone marrow mesenchymal stem cells obtained by these methods gradually lose their ability to proliferate and differentiate during subculture.

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