摘要
目的对大鼠骨髓间充质干细胞(bone marrow mesenchymal stem cells,BMSCs)进行分离培养与鉴定,并使其诱导分化为神经细胞。方法采用密度梯度离心和贴壁培养相结合的方法分离培养BMSCs,观察细胞形态变化及生长、增殖情况;对细胞进行表面标志物CD14、CD34、CD44、CD29免疫荧光染色及向骨、软骨和脂肪分化能力的鉴定;选用第3代细胞,经全反式维甲酸(retinoic acid,RA)和脑源性神经生长因子(brain-derived neurotrophy factor,BDNF)联合诱导后,免疫荧光染色检测神经前体细胞标记物Nestin及神经细胞标记物NSE的表达情况。结果体外培养的BMSCs呈成纤维细胞样,CD44和CD29的阳性率在90%以上,具有明显的向骨、软骨和脂肪分化的能力,经诱导后可分化为神经细胞。结论成功建立了体外分离扩增BMSCs的培养体系,所获得的细胞纯度高、生物学特征稳定,并可在体外诱导分化为神经细胞,从而为下一步细胞移植治疗脑缺血动物模型提供种子细胞。
Objective To isolate and culture the rat bone marrow mesenchymal stem cells（BMSCs） and to induce them into neural cells.Methods BMSCs were isolated with density gradient centrifugation and cell adhesion from rat bone marrow.The cell morphology,growth,proliferating were investigated.The surface markers CD14、CD34、CD44、CD29 were analyzed by immunofluocence.Osteogenic,chondrogenic,adipogenic differentiation potential were detected as well.At the 3rd passage,BMSCs were exposed to neural induction medium supplied with RA and BDNF,the induced samples were stained for Nestin and NSE detection by immunofluocence.Results Cultured BMSCs like fibroblast-shaped in vitro,the positive ratio of CD44 and CD29 were all above 90%,the BMSCs had potentials to differentiate into bone,cartilage,adipose and neural cells.Conclusion The culture system for BMSCs has been established,the cells have high purity and stable biological characteristics,they are differentiated into neural cells efficiently,and will be used to treat animal model of cerebral ischemia as seed cells.
出处
《哈尔滨医科大学学报》
CAS
北大核心
2011年第2期109-112,共4页
Journal of Harbin Medical University
基金
黑龙江省教育厅科学技术研究基金资助项目(11511162)
关键词
骨髓间充质干细胞
培养
诱导
神经细胞
bone marrow mesenchymal stem cells
culture
induce
neural cells
