江苏大学学报（医学版）2013，Vol.23Issue(3)：185-190,6.
人脂肪间充质干细胞的原代培养及体外成骨成脂诱导分化
Osteogenic differentiation of primary cultured human adipose-derived mesenchymal stem cells accompanying adipogenic differentiation in vitro
摘要
Abstract
Objective:To establish the isolation and culture method of human adipose-derived mesenchymal stem cells (hADSCs) derived from human adipose in vitro,so as to explore their morphology,identify cell surface markers,observe biological properties,and discuss the possible causes and mechanism of the phenomenon of hADSCs' osteogenic differentiation accompanying with synthesis of triglycerides.Methods:Human adipose tissue were obtained from abdominal operation.The hADSCs were isolated from human adipose tissue by 0.15％ collagenase digesting.The cells were applied to do the experiments:MTT method,flow cytometry,immunofluorescence.Its differentiation potential was proved by osteogenic and adipogenic differentiation.The osteogenic and adipogenic related genes:PPARγ-2,osteopontin expression were detected by real-time fluorescent quantitative PCR technique.Results:After isolation and culture,we obtained a large amount of hADSCs,which grew like swirls.MTT revealed high capability for and proliferation.The flow cytometry showed CD29 +,CD31-,CD34-,CD45-,CD73 +,CD105 +,CD166 +,HLA-DR-,which fit the results of immuno fluorescence.Moreover,these cells could be functionally induced into adipocytes and osteoblasts in the presence of appropriate conditioned media.During osteogenic differentiation,we found it accompanying with the synthesis of triglycerides.RT-PCR results proved that during the differention process,osteogenic and adipogenic related genes began to be expressed gradually,which had statistically significant(P ＜0.01).Conclusion:Highly efficient isolation and cultivation methods for hADSCs have been developed.They are a kind of mesenchymal cells with great application prospect,which characterized with adherent growth,high proliferation,stem cell phenotype and multipotent differentiation.During vitro osteogenic differentiation,the triglyceride formation has a certain role in promoting osteogenesis.关键词
人脂肪间充质干细胞/鉴定/成骨分化/成脂分化Key words
human adipose derived stem cells/ identify/ osteoblastic differentiation/ adipose differentiation分类
医药卫生引用本文复制引用
闵敏,张雪静,马红,许辉,李遇梅..人脂肪间充质干细胞的原代培养及体外成骨成脂诱导分化[J].江苏大学学报（医学版）,2013,23(3):185-190,6.基金项目
国家自然科学基金资助项目(30972663) （30972663）
