摘要
目的分离和培养大鼠骨髓间充质干细胞(marrow mesenchymal stem cells,MSCs),并行多向分化潜能的鉴定。方法经Percoll梯度分离法获得大鼠骨髓单个核细胞。贴壁培养后,通过倒置显微镜进行细胞形态学观察,并采用流式细胞术进行分析。传至第3代后,分别进行成脂肪细胞、神经元样细胞、血管内皮样细胞的诱导,并行免疫组织化学鉴定。结果骨髓间充质干细胞大小较为均匀,基本上呈梭形或星形的上皮样细胞,传代培养后的细胞体积增大,成纤维样细胞逐渐增多。经反复传代纯化的MSCs检测CD71、CD29呈阳性表达并证实骨髓间充质干细胞经诱导后分化为脂肪细胞、神经元样细胞、血管内皮样细胞。结论在体外,大鼠骨髓间充质干细胞具有多向分化潜能,可作为组织工程的种子细胞。
Objective To isolate and culture mesenchymal stem cells （MSCs） from bone marrow of adult rats and to identify the multilineage differentiation potential of the cells. Methods The mononuclear cells were obtained after Percoll gradient centrifugation. After adherence MSCs were cultured, the morphologic changes of MSCs were detected by inverted light microscopy. The expressions of CD71, CD29, CD34 and CD45 in MSCs were detected and identified by flow cytometry. The third passage cells were harvested and induced to differentiate into adipose cells, neuron-like cells, and vascular endothelium-like cells, then they were identified by immunohistochemical staining respectively. Results The results showed that the cultured MSCs were generally spindle-shaped or star-shaped epidermal-liked cells. The cellular volume increased and fibroblast-like cells were increased after serial subcultivation. After several generations they turned out to be positive expressions of CD29, CD71. After induction, they were confirmed to be adipose cells, neuron-like cells and vascular endothelium like cells by immunohistochemical staining. Conclusions In vitro MSCs have multilineage differentiation potential and can be used as the seed cells of tissue engineering.
出处
《复旦学报（医学版）》
CAS
CSCD
北大核心
2009年第5期528-532,共5页
Fudan University Journal of Medical Sciences
基金
国家自然科学基金项目(30560159)
教育部"新世纪优秀人才支持计划"(NCET-05-0820)项目
贵州省科技攻关项目基金(黔科字2008-7号)
关键词
骨髓间充质干细胞
脂肪细胞
神经元样细胞
血管内皮样细胞
marrow mesenchymal stem cell
adipose cells
neuron-like cells
vascular endothelium like cells
