High-efficiency method for inducing human mesenchymal stem cells to differentiate into cartilage

Abstract

The invention discloses a method for inducing human mesenchymal stem cells to differentiate into cartilage in vitro. The method comprises the following steps: (1) preparing a cell culture medium, and after preparation, preserving at 2-8 DEG C for later use; (2) preparing a compound cartilage induced differentiation culture medium, and after preparation, storing at 2-8 DEG C for later use; (3) resuspending the mesenchymal stem cells by using a cell culture medium, adjusting the density of the mesenchymal stem cells, and inoculating the mesenchymal stem cells by using a 96-well plate; (4) after the inoculation is finished, placing the culture medium in a CO2 incubator, standing for 2-3 hours, and supplementing a cell culture medium for subsequent culture; and (5) after standing for 24 hours, replacing the culture medium with a cartilage induced differentiation culture medium, and carrying out cartilage induced differentiation. The method has good stem cell culture and protection effects, stem cells obtained after cartilage differentiation are all pellet-shaped and regular in shape, obvious stem cell damage is avoided, the cartilage time period efficiency is high, and the method is suitable for being popularized in the field of stem cells and has wide development prospects.

Description

technical field [0001] The invention relates to the field of stem cells, in particular to a method for inducing differentiation of human mesenchymal stem cells into cartilage with high efficiency. Background technique [0002] The differentiation of stem cells is the selective activation or differential expression of some genes, thereby controlling the cell phenotype and the specific distribution of proteins. The differentiation of mesenchymal stem cells into specific cell types mainly depends on the activation of genes, and the types and concentrations of various factors in the extracellular microenvironment are important factors for gene activation. At present, the methods of inducing differentiation of mesenchymal stem cells into chondrocytes in vitro mainly include: (1) traditional chemical inducer method; (2) growth factor induction method; (3) combination of growth factor and chemical inducer; (4) co-culture Or use the conditioned medium close to the physiological sta...

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Problems solved by technology

However, this application adds fetal bovine serum albumin to the cell culture medium cultured in vitro, and the biological activity of fetal bovine serum is relatively high. Higher activity, it is easy to promote stem cells in the process of induction and differentiation, and the shape of cell pellets after cell differentiation is irregular or not into pellets; and the faster induction of differentiation into cartilage is likely to reduce the quality of stem cells differentiated into cartilage In addition, during the use of fetal bovine serum, due to its complex internal components and being easily affected by some external factors, it is easy to adversely affect the quality of the medium, such as containing viruses, which seriously affects the proliferation and cultivation of stem cells. The effect of chondrogenic differentiation

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