The application of BMP - Smad signaling pathway in the optimization of directed differentiation of murine embryonic stem cells into osteoblasts in vitro
XIA Quan
BAO Qian
JIANG De-ju
GAO Wei
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.
Keywords：murine embryonic stem cellsBMP-Smad signalingdifferentiationosteoblast
Publication Date：2018-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：6( 822-827 )