The study for induction and differentiation of rat's adipose-derived stem cells by valproic acid in vitro
10.3760/cma.j.issn.1001-2036.2018.04.010
- VernacularTitle:丙戊酸诱导鼠脂肪源性干细胞向许旺细胞分化的实验研究
- Author:
Fei WU
1
;
Ting JIA
;
Yue YANG
;
Zhibo SUN
;
Feng LIU
;
Xiangyang ZHANG
Author Information
1. 武汉大学人民医院骨外科
- Keywords:
Adipose-derived stem cells;
Schwann cells;
Valproic acid;
Induction and differentiation
- From:
Chinese Journal of Microsurgery
2018;41(4):356-360
- CountryChina
- Language:Chinese
-
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 AD-SCs 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),but no significant differ ence was found between groups 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),and groups B and D showed significant difference (P＜0.05).However,the expression of Nestin showed no significant difference among these groups (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.
