乳牙牙髓干细胞体外诱导成骨细胞的实验研究

A study of osteogenic induction of dental pulp stem cells from deciduous teeth in vitro

目的:体外分离、培养人乳牙牙髓干细胞,并向成骨细胞诱导。方法:采用酶消化法分离获得人乳牙牙髓干细胞,有限稀释法分离纯化,测定细胞克隆形成率,细胞计数法测定生长曲线,细胞爬片行HE染色,抗波形蛋白(Vimentin)、CD44和STRO-1免疫组化染色,并向成骨细胞诱导,行HE染色、碱性磷酸酶染色、Von Kossa染色、Van Gieson染色和抗骨钙素免疫组化染色进行鉴定。结果:通过有限稀释法获得了人乳牙牙髓干细胞,并诱导成成骨细胞,表现出与典型成骨细胞相似的形态特征和生物学特征。结论:成功从人乳牙牙髓中分离出牙髓干细胞,并诱导为成骨细胞。

PURPOSE： To isolate and culture dental pulp stem cells from human deciduous teeth, and induce them to osteoblasts, METHODS： Dental pulp stem cells were separated from deciduous teeth with enzyme digestion,then isolated and purified by limited dilution. The cells clone form rate was carried out, and cell growth curve was measured by count- ing the number of cells, HE stain and immunhistochemistry stain of Vimentin,CD44 and STRO-1 were tested. Then the cells were induced to osteoblasts, and HE stain,AKP stain, Von Kossa stain,Van Gieson stain,and immunhistochemistry stain of osteocalcin were used for evaluation, RESULTS ： Dental pulp stem cells were obtained from deciduous teeth by limited dilution. The cells were induced to osteoblasts, which elicited a biological and morphologic characteristics similar to those of osteoblasts, CONCLUSIONS ： The results show that dental pulp stem cells are obtained from deciduous teeth and induced to osteoblasts successfully.