胚胎面突外胚间充质干细胞向成骨细胞分化的实验研究

INDUCED DIFFERENTIATION OF ECTOMESENCHYMAL STEM CELLS OF HUMAN EMBRYO FACIAL PROCESS INTO OSTEOBLAST IN VITRO

OBJECTIVE: To investigate the possibility of ectomesenchymal stem cell of human embryo facial process in differentiating into osteoblasts. METHODS: Ectomesenchymal stem cells of human embryo facial process were isolated and cultured in mineralized promoting solution containing 10 mmol/L beta-glycerophosphate, 100 microg/ml ascorbic acid and 10 nmol/L dexamethasone supplemented with 15% FBS. The morphological change was observed by phase contrast microscopy. The characteristics of cells was identified by immunohistochemistry assay. Alkaline phosphatase activity was tested and the form of mineralized nodules was tested with Von Kossa staining. The expression of osteocalcin was identified by RT-PCR. RESULTS: There were significant changes in the shape of the cells after 3 days cultured in mineralized promoting solution. The cells became larger and the shape changed from fibroblast-like to multilateral. The result for anti-collogen type I staining was positive. The alkaline phosphatase activity increased. Mineralized nodules were formed after cultured 25 days by Von Kossa staining. RT-PCR assay showed induced cells expressed osteocalcin. CONCLUSION: Ectomesenchymal stem cells of human embryo facial process can be induced to differentiate into osteoblasts by mineralized promoting solution.