摘要
该研究探讨从颞下颌关节骨关节病患者的关节冲洗液中获取滑膜间充质干细胞(synovial mesenchymal stem cells,SMSCs)并进行体外培养,研究其间充质干细胞特性。采用离心、贴壁培养法从颞下颌关节骨关节病患者的关节冲洗液中提取并分离细胞,进行细胞形态学观察、细胞生长及周期的分析,细胞表面物标志物流式细胞术检测,间充质干细胞标志物免疫荧光检测,成骨、成脂、成软骨多向分化能力检测,促血管分泌因子检测。结果显示,通过离心、贴壁培养法可稳定地从患者颞下颌关节冲洗液中获取滑膜间充质干细胞,体外培养呈对数生长。SMSCs表达间质源性细胞的特异性标记物CD73、CD90、CD105、波形蛋白(Vimentin)以及α平滑肌肌动蛋白的特异性标记物α-SMA。经诱导培养基诱导,SMSCs可分化为骨、软骨和脂肪细胞,细胞分泌液中可检测到如成纤维细胞生长因子(fibroblast growth factor,FGF)、angiopoietin、转化生长因子-β(transforming growth factor-β,TGF-β)等10余种促血管生长因子。以上结果表明,该研究成功建立了以微创的方式获取颞下颌关节骨关节病患者的滑膜间充质干细胞,有望为组织工程应用于骨软骨修复提供良好种子细胞的来源。
Our objective is to isolate and identify the character of synovial mesenchymal stem cells（SMSCs）derived from the rinse solution of patients with temporomandibular joint osteoarthritis.Synovial-derived mesenchymal stem cells were collected from rinse solution of the temporomandibular joint osteoarthritis patients,cultured in vitro and the cell growth curve and cell morphology were analyzed;cell surface markers were detected by immunofluorescence and stem cell markers were detected by flow cytometry.We also detected the osteogenic,adipogenic and chondrogenic differentiation ability and the secretion factor of the cell.The results showed that we could steadily obtain the SMSCs from rinse solution of the temporomandibular joint osteoarthritis patients by means of centrifugation and adherent culture.Our results showed the logarithmic growth in vitro,expression of the surface markers（CD73,CD90,CD105）,vimentin and alpha-SMA in cultured SMSCs.The SMSCs could also be induced to differentiate into osteogenic,chondrogenic and adipogenic lineages in vitro.More than 10 kinds of vascular growth factors such as FGF,angiopoietin,TGF-βcan be detected from the cell secretion.These results indicated that SMSCs were obtained by minimally invasive method which provided a new resource of seed cell for the treatment of temporomandibular joint osteoarthritis.
出处
《中国细胞生物学学报》
CAS
CSCD
2017年第10期1312-1319,共8页
Chinese Journal of Cell Biology
基金
重庆市社会民生科技创新专项(批准号:cstc2016shmzx00010)
2016年重庆高校创新团队建设计划资助项目
重庆市高校市级口腔生物医学工程重点实验室资助项目资助的课题~~
关键词
颞下颌关节骨关节炎
间充质干细胞
滑膜
关节液
修复
temporomandibular osteoarthritis
mesenchymal stem cell
synovial membrane
joint synovia
renovation
