广东医学2018，Vol.39Issue(6)：822-827,6.
BMP-Smad信号通路在小鼠胚胎干细胞诱导分化成骨细胞方案优化中的应用
The application of BMP - Smad signaling pathway in the optimization of directed differentiation of murine embryonic stem cells into osteoblasts in vitro
摘要
Abstract
Objective To screen out the stable plan with the highest ratio of directed differentiation of murine embryonic stem cells (MSCs) into osteoblasts in vitro using BMP - Smad signaling pathway. Methods Murine MSCs were obtained via suspension culture form embryoid bodies (EBs). On the 14th to 21st day, the cells were assigned into 4 groups, Group A (culture supplemented with 50 μg/mL ascorbic acid + 50mmol/L β - glycerophosphate + 10-8 mol/ L dexamethasone), Group B (50 μg/mL ascormc acid + 10 mmol/L β - glycerophosphate + 1 μmol/L dexamethasone), Group C (50 μg/mL ascorbic acid + 50 mmol/L β - glycerophosphate + 1 μmol/L dexamethasone), and Group D (no inducer, as control group). The alkaline phosphonate (ALP) acivity was assessed on Day 7, 14, 21 and 28, On Day 22, the osteoblasts were determined by the 1 ％ Alizarin red staining. The expressions of BMP2, BMPR2, Smad1 and Smad5 were determined by real time - PCR and western blot. Results Compared with the control group, the activities of ALP in the other 3 groups were significantly increased from Day 7 after adherence, and reached the peak on Day 21. After Alizarin red staining, orange mineralized nodules of varying size was observed in osteoblasts. The osteoblast ratio in Group A, B, C and D were 15.34％, 23.71％, 29.12％ and 0.66％, respectively. Although the expression of BMP2, BM-PR2, Smad1 and Smad5 were increased or reduced in Group A and B, when compared with Group D, they were all significantly increased in Group C (P＜0.05). Conclusion The osteoblast ratio in Group C is the highest, suggesting the optimal osteoblast induction.关键词
小鼠胚胎干细胞/BMP-Smad信号通路/诱导分化/成骨细胞Key words
murine embryonic stem cells/BMP-Smad signaling/differentiation/osteoblast引用本文复制引用
夏荃,鲍倩,蒋德菊,高巍..BMP-Smad信号通路在小鼠胚胎干细胞诱导分化成骨细胞方案优化中的应用[J].广东医学,2018,39(6):822-827,6.基金项目
广东省自然科学基金资助项目(编号:2014A030313414) （编号:2014A030313414）
