颞下颌关节滑膜间充质干细胞成骨潜能的实验研究
Osteogenic Differentiation of Synovial Mesenchymal Stem Cellsin vitro
作者：李健,龙星,朱帆,杨雪超
Author：LI Jian1, LONG Xing1,ZHU Fan2, YANG Xue- chao3
收稿日期：2005-04-25 年卷（期）页码：2005,23(02):145-
期刊名称：华西口腔医学杂志
Journal Name：West China Journal of Stomatology
关键字：滑膜,间充质干细胞,成骨细胞,
Key words：synovial membrane,mesenchymal stem cells,osteogenesis,
基金项目：国家自然科学基金资助项目(30371549)
中文摘要
目的 研究滑膜间充质干细胞的成骨潜能。方法 用2 g/L的Ⅰ型胶原酶消化获得滑膜细胞,待细胞汇
合后用有限稀释法进行单细胞克隆,筛选出滑膜间充质干细胞(SMSC)。取第3代SMSC进行成骨诱导培养,每天观
察细胞形态,并于培养7 d后检测ALP活性和骨桥蛋白的表达,RT-PCR检测核心结合因子al(cbfal)mRNA的转录;
培养30 d后作Von Kossa′s染色,检测成骨化程度。结果 SMSC在体外培养形成葵花样细胞集落,胞浆突起明显,
相互连接成网状;成骨化诱导剂可诱导SMSC定向分化为多形性成骨细胞,细胞ALP染色阳性,骨桥蛋白强阳性,电
泳显示有cbfal的特异性条带,矿化区经Von Kossa′s染色呈阳性反应。结论 经体外纯化的SMSC可定向诱导分化
为成骨细胞,符合成骨细胞的生物学特征。
英文摘要
Objective To investigate the potential of synovial mesenchymal stem cells(SMSC) in osteogenic differentiation. Methods SMSCwere obtained by limited dilutionmethod and expanded to culture in 25-milliliterflasks. The attached cellswere treated with inductive medium containing dexamethasone, glycerophosphate and vitamin C at 3rd passage SMSC. The mineralized nodule was stained byVonKossa method. The expression ofALP and osteopontinwere detected by histochemical, immunohistolog- ical staining technique, respectively, while the expression of cbfa1 mRNAby RT-PCR.Results Pure SMSCwhichwere of spin- dle shape and star shape, uniform in size, could be induced to pleomorphism osteoblastin vitro, whichwere intensive positive in ALP and osteopontin. The expression of cbfa1 mRNAwere also verified by RT-PCR and the polygonal cells formed nodular struc- ture at 4 weeks. All these were coincidentwith the characters of osteoblast.Conclusion SMSC can be purified and induced into osteoblastin vitro.
【关闭】
