Study On Differentiations Of MSCs Into Neuron-like Cells Induced By Herba Cynomorii Serum

Objective: This research aimed to establish the methods of separating, culture and purifying rat mesenchymal stem cells(MSCs) in vitro, and searched after the ability of directive differentiation that the MSCs were induced into neuron-like cells by Herba Cynomorii (HC) Serum, which provided the initial experimental foundation for the cells alternative therapies of MSCs.Methods: Under sterile conditions MSCs were collected from the femur of the SD rats as primary culture, then cultured, purified and expended in vitro according to ability of adherent culture. According to ability of adherence, sieve easily adherent but not sbatle cells to subculture. The cells shape and growth were observed by inverted microscope day-to-day. Different doses of HC serum were used to induce MSCs into neuron-like cells in vitro when the third generation of cells were developed. Meanwhile, the change of cells shape and quantity also need to be observed when it splited. Then detected the neural stem cells markers, such as Nestin, neuron specific enolase (NSE) and glial fibrillary acidic protein (GFAP) by immunocytochemistry, and calculate the rate of protein conversion.Results:â‘ MSCs were separated successfully and amplified in a large amount by adhesion culture in vitro.â‘¡HC serum can directly induce MSCs differentiation into typical neuron-like cells, meanwhile, Nestin and NSE have higher expression, expect GFAP.â‘¢The effect of HC serum inducing MSCs into neuron-like cells was better than negtive control group (P<0.05) , but less than 6-Mercaptoethanol group (P<0.05).â‘£There was evident difference of the ability of inducing MSCs into neuron-like cells between low-dose and high-dose HC serum groups (P<0.05) , but there were no significant difference either between low-dose and moderate-dose HC serum groups (P>0.05) , or between moderate-dose and high-dose HC serum groups (P>0.05). Conclusions: MSCs can be separated and cultured from bone marrow of SD rats in vitro according to the adhesive ability. HC serum can directly induce MSCs differentiation intotypical neuron-like cells.