改良组织块酶消化法原代培养人牙髓干细胞的研究

Primary culture of hPDSCs with modified enzymatic digestion-explants method

目的探索应用改良组织块酶消化法原代培养人牙髓干细胞并对细胞进行鉴定。方法选取18~26岁健康人因正畸或阻生而拔除的健康、完整、无龋坏的牙齿30颗,采用改良组织块酶消化法原代培养人牙髓干细胞并通过对细胞的形态观察、细胞表面标记物检测、多向分化能力检测和生长曲线检测等方法鉴定人牙髓干细胞。结果对人牙髓组织经改良组织块酶消化法原代培养得到的细胞形态类似于成纤维细胞和间充质细胞,生长曲线为S型,细胞表面标记物检测显示间充质干细胞标记物CD29、CD44、CD90表达阳性,造血干细胞表面标记物CD34、CD45、CD106表达阴性,同时细胞具有多向细胞分化的能力。结论应用改良组织块酶消化法可以从人牙髓组织中成功原代培养人牙髓干细胞。

Objective To explore methods using modified tissue enzymatic separations for culturing primary hDPSCs in vitro and further identify the cells produced.Methods Primary hDPSCs were cultured using the modified tissue enzymatic separation method,and cells were identified by morphology,cell surface markers,and differentiation potential and evaluated using flow cytometry and growth curves.Results The hDPSCs were successfully isolated using the modified tissue enzymatic separation method.The morphology of these cells was similar to that of fibroblasts and mesenchymal stem cells,and the growth curve was"S"-type.The results of cell phenotype analysis indicated that the cells were positive for surface markers of mesenchymal stem cells,including CD29,CD44,and CD90,and negative for markers of hematopoietic stem cells,including CD34,CD45,and CD106.The cells were capable of differentiating intomultiple cell types.Conclusion The modified tissue enzymatic separation method can successful be used to culture primary hDPSCs in vitro.