Culture solution for inducing mesenchymal stem cells to differentiate into islet-like cells, and inducing method and application of culture solution

Abstract

The invention discloses a culture solution for inducing mesenchymal stem cells to differentiate into islet-like cells, and an inducing method and application of the culture solution. The culture solution comprises the following materials: niacinamide, Conophylline, cell growth factor, betacellulin and a base medium. The base medium contains 97% of high glucose DMEM (Dulbecco Modified Eagle Medium), 2% of B-27 and 1% of N-2. The inducing method comprises the following steps of: preparing an inducing culture solution; preparing the human mesenchymal stem cells; taking the human mesenchymal stem cells, inoculating the human mesenchymal stem cells into a six-hole ultralow absorption culture plate by 1.5-2*105cells / hole, adding 3ml inducing culture medium into each hole and carrying out suspended induction; and changing liquid at every 3 days, collecting cell supernatant at the ninth day, and storing the cell supernatant at the temperature of -20 DEG C. The culture solution disclosed by the invention has the advantages that the human mesenchymal stem cells are induced to differentiate into the islet-like cells by utilizing the combination of the niacinamide and the Conophylline, so that the inducing cycle is shortened, the suspension cells are beneficial to being clustered to form cell clusters similar to natural islets, further the induced differentiation efficiency is obviously increased and the clinical application risk is reduced; and the function of inducing the secretion of the cell insulin is obviously improved.

Description

technical field [0001] The present invention relates to a culture medium, an induction method and an application for efficiently inducing human adipose stem cells to differentiate into functional islet-like cell clusters by using the novel plant inducer Conophylline and nicotinamide. Background technique [0002] Type 1 diabetes, also known as insulin-dependent diabetes, is an autoimmune disease mediated by primary T cells, which eventually leads to massive destruction of β cells, insufficient insulin secretion, and unbalanced regulation of blood sugar concentration, seriously endangering human health. Islet transplantation is the most effective way to treat diabetes, but there is a serious shortage of donor cells, so it is necessary to find new substitutes for islet transplantation. Mesenchymal stem cells are a kind of cells with multi-lineage differentiation potential, which can be directional induced into various cells such as islet-like cells and chondrocytes. Moreover,...

AI Technical Summary

Problems solved by technology

Due to the strong adherence of mesenchymal stem cells, it hinders the aggregation of cells during the induction process, so the induction rate is generally low

The two-part method or the three-step method uses more induction factors, and the induction period is generally 14-21 days. The induction period is long and there are many types of residual factors, which increases the risk of clinical application.

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