Culture solution and method for inducing mesenchymal stem cells to differentiate into glomerular mesangial cells

Abstract

The invention discloses culture solution for inducing mesenchymal stem cells to differentiate into glomerular mesangial cells and an inducing method. The culture solution comprises a human platelet-derived growth factor-BB, all-trans-retinoic acid, collagen-IV and a basic medium, wherein the basic medium is a low-sugar DMEM (Dulbecco's modified eagle medium) containing 2% of horse serum and 5ng / ml of bFGF (basic fibroblast growth factor). The inducing method includes the steps: preparing the inducing culture solution; preparing the mesenchymal stem cells; coating a culture plate by the collagen-IV with the concentration of 5ug / cm<2>; inoculating the mesenchymal stem cells with the cell density of 5*103 / cm<2> into the coated culture plate and adding the inducing culture solution; changing the solution once every three days and performing inducing culture for seven days. The mesenchymal stem cells are jointly induced to differentiate into the glomerular mesangial cells by the aid of the human platelet-derived growth factor-BB, the all-trans-retinoic acid and the collagen-IV, inducing period is short, inducing efficiency is high, the glomerular mesangial cells formed by differentiation are high in functional activity, and transplantation therapy of kidney diseases such as kidney failure is facilitated.

Description

technical field [0001] The invention relates to a culture medium and an induction method for efficiently inducing the differentiation of mesenchymal stem cells into glomerular mesangial cells by using the combination of novel cell growth factor human platelet-derived growth factor-BB and all-trans retinoic acid. Background technique [0002] When various kidney diseases gradually develop into end-stage renal failure, they can usually only be treated with the help of renal replacement means, mainly dialysis and kidney transplantation. However, there are obstacles and deficiencies that are difficult to overcome in these two treatment methods. Among them, dialysis is limited to the replacement of filtration function, and does not have a series of important functions such as metabolism, secretion and maintenance of internal environment homeostasis of normal kidney. Although kidney transplantation has better therapeutic effect than dialysis, it also faces immune rejection, Due t...

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Problems solved by technology

[0004] The present invention establishes a new method for inducing mesenchymal stem cells to differentiate into glomerular mesangial cells, using platelet-derived growth factor-BB and all-trans retinoic acid in combination, and at the same time supplementing type IV collagen as the extracellular matrix, Compared with the current commonly used induction methods, the induction efficiency and induction cycle are improved, and the induced glomerular mesangial precursor cells obtained have strong biological activity, which is helpful for better application in clinical transplantation, overcoming the The shortcomings of the existing inducers and induction methods such as long induction period, low efficiency, and poor function

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