人牙周膜干细胞的体外分离、纯化及初步鉴定  

Isolation,purification and identification of human periodontal ligament stem cells

作　　者：高秦[1] 刘宏伟[2] 金岩[3] 聂鑫[3] 刘源[3] 

机构地区：[1]广州南方医科大学南方医院,510515 [2]上海同济大学口腔医学院 [3]西安第四军医大学口腔医院病理科

出　　处：《实用口腔医学杂志》2006年第1期34-37,共4页Journal of Practical Stomatology

基　　金：陕西省自然科学研究计划;2003C2042

摘　　要：目的:体外分离纯化人牙周膜干细胞,研究其生物学特性及表型特点并进行初步鉴定。方法:采用胶原酶和D ispase酶联合消化法培养人牙周膜干细胞,有限稀释法分离纯化,将克隆形成的细胞通过透射电镜观察其超微结构、流式细胞仪细胞周期分析及免疫组织化学染色技术检测抗波形丝蛋白(V im en-tin)、STRO-1、骨粘素(ON)、骨涎蛋白(BSP)的表达。结果:获得纯化人牙周膜干细胞,在透射电镜下,这种细胞核质比例大,核大,细胞器少;流式细胞仪细胞周期分析大多数细胞处于G0/G1期,为慢周期性;该细胞抗波形丝蛋白、STRO-1表达阳性、骨粘连素、骨桥蛋白弱阳性表达。结论:提供了比较有效的分离纯化牙周膜干细胞的方法。该方法分离的人牙周膜干细胞具有干细胞的超微结构、细胞周期及表型特点。Objective：To isolate and identify the periodontal ligament stem cells. Methods：Periodontal ligament tissues were digested with a mixture of collagenase and Dispase, then the periodontal ligament stem cell clones were isolated by limited dilution method. The transmission electron microscopy （TEM） , flow cytometery and immunobistocbemistry procedure were employed to study the uhrastructure, cell cycle and surface marker of the cloned cells. Results ：The obtained cells showed the characteristics of undifferentiated morphology at uhrastructaral level. 94, 4% of the cells were in phase G0/G1. The cells were Vimentin, STRO-1, ON and BSP positive. Conclusion：Cloning incubation may be the effective way to isolate and purify the periodontal ligament stem cells.

关 键 词：人牙周膜干细胞 细胞克隆 超微结构 细胞周期 免疫细胞化学 纯化技术 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]