Culture method for inducing chondrogenic differentiation of adipose-derived stem cells and culture solution

Abstract

The invention discloses a culture method for inducing chondrogenic differentiation of adipose-derived stem cells and a culture solution. The culture method comprises the following steps: (1) taking asubcutaneous fatty tissue, washing the subcutaneous fatty tissue, removing an adipose tissue membrane and blood vessels and shearing the subcutaneous fatty tissue into broken blocks, carrying out enzymic digestion, adding a complete medium, centrifuging to remove adipose cells and lipid droplets, resuspending on the complete medium, inoculating into a culture bottle, culturing for 24 h and then changing the culture solution to remove cells which are not attached to walls, changing the solution every three days, digesting for 3-5 min with 0.25% of pancreatin and 1 mmol / L EDTA at the temperatureof 37 DEG C after cell fusion, stopping digestion of the complete medium, and carrying out passage at the ratio of 1: 2; and (2) inoculating passage cells into the culture bottle which contains the complete medium, culturing for 48h and then changing the medium into an incomplete medium, adding a Sox9 gene activator to continue culturing, and changing the medium every three days. By the providedmethod, chondrogenic differentiation of the adipose-derived stem cells can be induced effectively, the adipose-derived stem cells can be used as seed cells to prepare a large number of chondrocytes tomeet clinic requirements to the chondrocytes.

Description

technical field [0001] The invention belongs to the field of biology, and relates to a culture method and a culture solution for inducing chondrogenic differentiation of adipose stem cells. Background technique [0002] At present, in the field of research on mesenchymal stem cells, the main target is bone marrow mesenchymal stem cells, and the method of isolation and culture has been initially formed, but the mass culture of bone marrow mesenchymal stem cells is limited. Especially in the study of cartilage tissue engineering, due to the difficulty of isolating and culturing bone marrow mesenchymal stem cells, it is limited to become a good source of tissue engineering seed cells. Therefore, recently people have turned their research to adipose-derived stem cells, and found that the cells have strong proliferation and differentiation capabilities, and are easier to isolate and culture. In clinical research, adipose-derived stem cells also have broad application prospects i...

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Problems solved by technology

[0002] At present, in the field of research on mesenchymal stem cells, the main target is bone marrow mesenchymal stem cells, and a method of isolation and culture has been initially formed, but there are certain limitations in cultivating bone marrow mesenchymal stem cells in large quantities.

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