Method for promoting chondrogenic differentiation of umbilical cord mesenchymal stem cells

Abstract

The invention provides a method for promoting chondrogenic differentiation of umbilical cord mesenchymal stem cells, which comprises the following steps: taking out the umbilical cord tissue, cutting it into pieces and cleaning it, adding tissue digestive juice for digestion, filtering and removing the supernatant after the digestion is terminated, and obtaining the umbilical cord Mesenchymal stem cells; the umbilical cord mesenchymal stem cells are cultivated with selective medium for umbilical cord mesenchymal stem cells; when the cells reach 80-90% confluence, the cells are digested with a low-toxicity digestive solution, and then the umbilical cord mesenchymal stem cells are used The cryopreservation solution was frozen and placed in liquid nitrogen; the cells were removed from the liquid nitrogen, quickly thawed in a 37°C water bath, centrifuged to remove the supernatant, and the cells were resuscitated by adding the cell resuscitation solution; the resuscitation solution was removed and the induction Culture in pre-activation solution; when the cells grow to 80-90% confluence, digest the cells with the low-toxicity digestion solution; inoculate the cells, culture overnight, remove the supernatant, add chondrogenic induction medium, and use 2% oxygen concentration cultured in a hypoxic incubator. Compared with the prior art, the method of the present invention can stably and effectively promote chondrogenic differentiation of umbilical cord mesenchymal stem cells with a shorter induction time.

Description

technical field [0001] The invention belongs to the field of stem cells and regenerative medicine, in particular to a method and culture medium for promoting chondrogenic differentiation of umbilical cord mesenchymal stem cells. Background technique [0002] Umbilical cord mesenchymal stem cells have multilineage differentiation ability. The differentiation effect of mesenchymal stem cells is mainly determined by the activity of stem cells and induction conditions. [0003] Mesenchymal stem cell chondrogenesis is a multi-step cellular event, which begins with the aggregation of a special subset of mesenchymal stem cells and then differentiates into cartilage. [0004] The commonly used method for obtaining umbilical cord mesenchymal stem cells is as follows: cut Wharton's jelly into pieces, and adhere to culture in complete medium (DMEM with 10% FBS) for 16 days. The conventional cryopreservation method is as follows: freeze in liquid nitrogen after programmed cooling in c...

AI Technical Summary

Problems solved by technology

[0005] The deficiencies of the above existing stem cell chondrogenic differentiation methods are as follows: the required induction time is longer; the use of support media to assist chondrogenic differentiation is more complicated and the cost is higher; although the cell viability will be detected before each induction, it cannot be guaranteed A batch of cells can complete chondrogenic differentiation, which will cause waste of cell resources

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