人脂肪间充质干细胞的分离及培养和鉴定

Isolation,culture and identification of human adipose-derived mesenchymal stem cells

目的:建立体外分离培养人脂肪间充质干细胞(hADSCs)的方法,观察其形态及生物学特性,探讨其作为颞骨上皮组织工程种子细胞的可行性。方法:整形外科吸脂术获得女性腹部皮下脂肪,0.3%Ⅰ型胶原酶消化,贴壁培养hADSCs,进行成脂、成骨诱导验证其多向分化潜能;流式细胞术检测细胞表面分子;MTT法绘制细胞增殖曲线。结果:该实验获得形态均一的长梭形、成纤维细胞样生长的hADSCs,具有成脂、成骨分化潜能;流式细胞术检测细胞高表达CD105、CD29、CD90,低表达CD34、CD45;细胞有明显的指数生长期。结论:该实验分离获得的细胞具有hADSCs特性,可作为颞骨上皮组织工程种子细胞。

Objective：To establish isolation and culture methods of human adipose derived mesenchymal stem cells（hADSCs）in vitro and to investigate the basic biological characteristics of hADSCs as seed cells involved in repair of epithelium during temporal bone surgery.Method：Human adipose tissue was collected after liposuction surgery from the cosmetic and plastic surgery center,and then was digested with 0.3% collagenase Ⅰ.hADSCs were isolated and adherent cultured.Multilineage differentiation potential of hADSCs were determined by differentiating cells into adipocytes and osteoblast.Phenotypes of cells were detected by flow cytometer,and cell growth curve was observed by MTT.Result：Obtained hADSCs were similar size,spindle-shaped fibroblasts-liked cells and had differentiation potential to adipocytes and osteoblast.Flow cytometry showed that cells were highly positive for CD105,CD90,CD29 and weakly positive for CD34 and CD45.MTT exhibited that hADSCs had a exponential growth phase.Conclusion：Isolated stem cells from human adipose tissue show the typical characterization of hADSCs,and can be used as seed cells involved in repair of epithelium during temporal bone surgery.