胎盘来源的间充质干细胞体外分离培养及向软骨细胞分化的研究

The Isolation,Culture and Differentiation of Human Placenta-derived Mesenchymal Stem Cells into Chondrocytes

目的对人胎盘间充质干细胞体外分离培养及向软骨细胞分化进行研究,为软骨组织工程提供种子细胞。方法取人正常分娩后的胎盘,用组织块培养法分离获取胎盘间充质干细胞,对所获细胞进行体外培养和表面标志的流式细胞仪分析。对经过证实的骨髓间充质干细胞用含TGF-β_3、维生素C、胰岛素、地塞米松、转铁蛋白的诱导培养基处理7-28d,对诱导细胞进行甲苯胺蓝染色及Ⅱ型胶原染色,鉴定诱导后的软骨细胞表型。结果培养的人胎盘间充质干细胞均一表达CD_(29)和CD_(44),而CD_(31)、CD_(34)、CD_(45)和HLA-DR呈阴性。细胞经诱导液作用14d后,甲苯胺蓝染色及Ⅱ型胶原染色阳性。结论采用组织块培养法能分离获取高纯度的人胎盘间充质干细胞,该细胞经诱导液作用可定向分化为软骨细胞。

Objective To study the isolation and culture and differentiation of human plaeenta-devived mesenchymal stem cell （HPDMSC） into chondrocytes as potential seed cells for cartilage tissue engineering. Methods Isolated HPDMSCs from human placenta were deliveried and cultured in Petri dish. Specific cell surface marks were identified by flow-cytometry. HPDMSCs were treated with inductive medium containing TGFβ3, vitamin C, insulin,dexathemesone and transferrin for 7-28 days. The toluidine blue staining and immunocytochemistry were peformed for chondrogenic phenotype identification. Results HPDMSCs expressed CD29 and CD44 positively, but CD34 and CD45 and HLA-DR negatively. After 14 days induction, the cells were positively stained by toluidine blue and expressed Ⅱ collagen positively. Conclusion The isolated HPDMSC can be amplified and induced to differentiate into chondrocytes in vitro by the present isolation and culture methods.