中文摘要：

目的分离和培养大鼠骨髓间充质干细胞（MSCs）,体外诱导分化为神经元样细胞及脂肪细胞。方法取SD大鼠股骨和胫骨骨髓,经贴壁筛选法体外分离、培养并纯化MSCs。通过倒置显微镜进行细胞形态学观察和流式细胞术进行分析。地塞米松和胰岛素诱导MSCs向脂肪细胞分化,采用油红O染色鉴定;在含碱性成纤维细胞生长因子（bFGF）,二甲基亚砜（DMSO）和3-叔丁基-4羟基茴香醚（BHA）培养基中诱导MSCs向神经元样细胞分化,通过免疫细胞化学法鉴定诱导后细胞神经元烯醇化酶（NSE）、神经巢蛋白（Nestin）、胶质纤维酸性蛋白（GFAP）的表达情况。结果倒置显微镜下观察到分离培养的骨髓间充质干细胞大小均匀,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。MSCs传至第3代采用流式细胞仪检测CD71、CD29呈阳性表达;经油红O染色证实,第5代MSCs经诱导后分化为脂肪细胞;而免疫细胞化学证实MSCs经诱导后分化为神经元样细胞,表达NSE、Nestin,不表达GFAP。结论在体外,大鼠MSCs可诱导分化为脂肪细胞、神经元样细胞,证明其具有多向分化潜能,是组织工程研究中最有前途的种子细胞之一。

英文摘要：

Objective To explore the cultivation of rat bone marrow mesenchymal stem cells （MSCs） and the potency of differentiating into adipose cells and neuron - like cells in vitro. Methods MSCs were obtained from bone marrow of SD rat＇s femoral and tibial bones ,cultured and propagated in vitro, then purified by adhesive - screening methods. The morphologic changes of MSCs were observed by inverted light microscope. The expressions of CD71 , CD29, CD34 and CD45 in MSCs were detected and identified by flow cytometry. The 5th passage MSCs were induced to differentiate into adipose cells in induction medium containing desamethasone and insulin. Then differentiated cells were identificated by oil red O stain. After treatment with basic fibrohlast factor（bFGF） , dimethyl suffoxide （DMSO） and butyl hydroxy anisole （BHA）, MSCs could be differentiated into cells with typical neuronal morphology. The expressions of neuron specifice nolase （ NSE ） , Nestin and glialfibriliary acidic protein（GFAP） were detected by immunocytochemistry. Results The results of inverted light microscope showed that the cultured MSCs appeared no difference, generally were spindle - shaped or star - shaped epidermal - liked cells. The cellular volume increased and fibroblast- like cells were increased after serial subcultivation. The 3th passage cells were detected by flow cytometry and the results turned out to be positive expressions of CD29, CD71. After induction,they were confirmed to be adipose cells by oil red O stain and neuron - like cells by immunocytochemistry staining, expressed NSE, Nestin, none of them expressed GFAP. Conclusions In vitro, MSCs can be induced into adipose cells and neuron -like cells ,which shows that MSCs have muhipotency and are one of the best seed cells in the tissue engineering.