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颞下颌关节滑膜间充质干细胞成骨潜能的实验研究
引用本文:李健,龙星,朱帆,杨雪超. 颞下颌关节滑膜间充质干细胞成骨潜能的实验研究[J]. 华西口腔医学杂志, 2005, 23(2): 145-147
作者姓名:李健  龙星  朱帆  杨雪超
作者单位:1.武汉大学口腔医院 口腔颌面外科;2.武汉大学生命科学学院;3.武汉大学口腔医学院 口腔生物医学工程教育部重点实验室,湖北 武汉430079
基金项目:国家自然科学基金资助项目(30371549)
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。

关 键 词:滑膜  间充质干细胞  成骨细胞  
文章编号:1000-1182(2005)02-0145-03
收稿时间:2005-04-25
修稿时间:2005-04-25

Osteogenic Differentiation of Synovial Mesenchymal Stem Cells in vitro
LI Jian,LONG Xing,ZHU Fan,YANG Xue-chao. Osteogenic Differentiation of Synovial Mesenchymal Stem Cells in vitro[J]. West China journal of stomatology, 2005, 23(2): 145-147
Authors:LI Jian  LONG Xing  ZHU Fan  YANG Xue-chao
Affiliation:1. Dept. ofOral and Maxillofacial Surgery, School andHospital ofStomatology, Wuhan University, Wuhan430079,China;2. Life Science Institute ofWuhan University, Wuhan430079, China;3. Key Lab. ofOral Biomedical Engineering ofMinistry ofEducation, Wuhan University, Wuhan430079, China
Abstract:Objective To investigate the potential of synovial mesenchymal stem cells(SMSC) in osteogenic differentiation.Methods SMSC were obtained by limited dilution method and expanded to culture in 25-milliliter flasks. The attached cells were treated with inductive medium containing dexamethasone, glycerophosphate and vitamin C at 3rd passage SMSC. The mineralized nodule was stained by Von Kossa method. The expression of ALP and osteopontin were detected by histochemical, immunohistological staining technique, respectively, while the expression of cbfa1 mRNA by RT-PCR.Results Pure SMSC which were of spindle shape and star shape, uniform in size, could be induced to pleomorphism osteoblast in vitro, which were intensive positive in ALP and osteopontin. The expression of cbfa1 mRNA were also verified by RT-PCR and the polygonal cells formed nodular structure at 4 weeks. All these were coincident with the characters of osteoblast.Conclusion SMSC can be purified and induced into osteoblast in vitro.
Keywords:synovial membrane  mesenchymal stem cells  osteogenesis
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