摘要
目的 体外定向诱导成人骨髓间质干细胞 (MSC)分化为神经元样细胞。方法 采用Ficoll Paque液 (10 77g/L)离心分离成人MSC ,体外扩增 ,分别采用含碱性成纤维细胞生长因子 (bFGF)和叔丁对甲氧酚 (BHA)或硫代甘油等试剂的无血清DMEM诱导MSC分化为神经元。免疫组化鉴定神经元烯醇化酶 (NSE)、神经丝蛋白 (NF)、胶质纤维酸性蛋白 (GFAP)、巢蛋白 (nestin)的表达。结果 成人骨髓间质干细胞在体外扩增原代可获得 5× 10 5,10代可获得 2× 10 10 个细胞。加入bFGF和BHA等诱导剂或硫代甘油诱导后 ,MSC胞体收缩 ,突起伸出 ;免疫组化显示诱导出的神经元样细胞NSE、NF、nestin表达阳性 ,GFAP阴性。结论 成人骨髓间质干细胞在体外可以分化为神经元样细胞。
Objective To investigate the differentiation from human mesenchymal stem cells (hMSC) into neuron-like cells. Methods hMSC were separated from rib marrow with Ficoll-Paque reagent and expanded in culture medium. hMSC were induced to differentiate into neurons with DMEM/BHA/DMSO or DMEM/monothioglycerol, respectively. Neuron-specific enolase (NSE), neurofilament (NF), nestin, glial fibrillary acidic protein (GFAP) were detected by immunohistochemistry. Results hMSC were expanded to be undifferentiated cells in culture for more than 10 passages. The isolated and cultured MSC comprised a single phenotypic population and displayed a fibroblast-like morphology. Simple method induced hMSC exhibiting a neuronal phenotype, with a positive expression of NSE, NF-M and nestin at 5 hours. But the neuron-like cells did not express the glial astrocyte marker GFAP. Conclusion It suggests that hMSC can be differentiated into neurons in vitro .
出处
《中华神经科杂志》
CAS
CSCD
北大核心
2002年第3期165-167,共3页
Chinese Journal of Neurology
基金
国家重点基础研究发展规划项目(973)资助 ( #G19990 5 430 0 )
