摘要
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.
作者
刘影
高杰
吴补领
LIU Ying;GAO Jie;WU Buling(Stomatological Hospital, Southern Medical University, Guangzhou 510280, China;Department of Stomatology, Guangzhou Medical University, Guangzhou 510182, China;Nanfang Hospital, Southern Medical University & Stomatological College of Southern Medical University, Guangzhou 510515, China)
出处
《口腔疾病防治》
2018年第3期166-170,共5页
Journal of Prevention and Treatment for Stomatological Diseases
基金
广东省自然科学基金项目(2015A030310071)
广东省医学科学技术研究基金项目(A2016212)
广东省自筹经费类科技计划项目(粤科规财字[2015]110号)
关键词
人牙髓干细胞
组织块酶消化法
原代培养
细胞鉴定
hDPSCs
Explants method and enzymatic separation method
Primary culture
Cells identification