The study for induction and differentiation of rat’s adipose-derived stem cells by valproic acid in vitro

Abstract

Objective To study the inducting differentiation effects of the vaproic acid (VPA) on rats adipose derived stem cells (ADSCs) in vitro. Methods From November, 2016 to October, 2017, the ADSCs were isolated from 2 healthy 3-weeks-old Sprague-Dawley (SD) rats and cultured to passage 3, which were treated with 10 ng/ml bFGF for 24 hours before induction. Then the induction media contained the VPA with different concentrations: group B (0.5 mmol/L, 200 μl) VPA; group C (1.0 mmol/L, 200 μl) VPA; group D (10 mmol/L, 200 μl) VPA, and D-Hank was used in group A as blank control group. The morphological changes of the cells were observed every day. At 7 days of induction, the gene expressions of neuron-specific enolase (NSE), nestin (NES), and S-100 were detected by real-time fluorescent quantitative PCR. The S-100 protein expression was tested by immunofluorescence staining. Significance of difference was determined using independent t test. Probabilities lower than 5% (P < 0.05) were considered statistically significant. Results At 4 days after induction, some ADSCs of groups B, C, and D showed the morphology of Schwann-like cells or neuron-like cells, the change of group C was more obvious; and the ADSCs of group A had no obvious change, which still present spindle. The S-100 immunofluorescence staining showed higher ratio of positive expression in groups B, C, and D (more obvious in group C) and lower ratio of positive expression in groups A and D (more obvious in group A). The gene expression of S-100 showed dose-dependent increases in groups C, which was significantly higher than that of groups A, B, and D (P 0.05). The gene expression of NSE showed the same tendency as S-100, which reached the peak in group C; the gene expression of NSE in group C was significantly higher than that of groups A, B, and D (P 0.05). Conclusion ADSCs can be induced to differentiate into Schwann-like cells or neuron-like cells under the treatment of VPA, and 1.0 mmol/L is the optimal concentration. Key words: Adipose-derived stem cells; Schwann cells; Valproic acid; Induction and differentiation