人牙周膜干细胞的分离培养及初步鉴定

Initial identification of human periodontal ligament stem cells

目的体外分离培养人类牙周膜干细胞并对其生物学性状进行初步鉴定。方法采用有限稀释法进行牙周膜干细胞克隆筛选,获得单细胞克隆来源细胞,检测其克隆形成率,并采用免疫组织化学染色、碱性磷酸酶染色、免疫荧光染色等方法鉴定牙周膜干细胞的组织来源及生物学特性。结果有限稀释法获得的牙周膜干细胞克隆株呈簇状生长,排列紧密,细胞呈圆形或椭圆形,细胞较小,排列形成紧密的团块状。免疫组化染色显示细胞波形蛋白阳性,角蛋白阴性;碱性磷酸酶染色阳性;早期间充质干细胞的标志物STRO-1荧光染色显示细胞发出明亮的红色荧光,表达阳性。结论牙周膜中存在具有高增殖能力的干细胞,克隆化分离培养的人牙周膜干细胞具有强克隆形成能力,具有间充质干细胞表型和生物学特性。

Objective To isolate, culture and investigate the characterizations of human periodontal ligament stem cells （hPDLSCs）. Methods Single-cell suspension was used to obtain hPLSCs clone, and the characterizations of hP- DLSCs were identified by immunohistochemistry staining, alkaline phosphate （ALP） staining and immunofluence staining. Results The colony of hPDLSCs could be obtained from single-cell suspension. The expressions of Vimentin and STRO-1 were positive; ALP staining showed strong staining. Conclusion hPDLSCs can he isolated and cultured from human periodontal ligament cells, hPDLSCs express biological marks of stem cells, and have similar characterizations with mesenchymal stem cells.