骨髓基质细胞体外诱导分化成神经元和胶质细胞

目的　探索骨髓基质细胞 (bonemarrowstromalcells,BMSC)体外诱导分化为神经元和神经胶质细胞的可行性 ,为BMSC在神经科学领域内的应用奠定基础。方法　以成年犬BMSC为实验对象 ,利用碱性成纤维细胞生长因子 (bFGF)、表皮生长因子 (EGF)、维甲酸 (RA)、脑源性神经营养因子 (BDNF)、胶质细胞系来源神经营养因子 (GDNF)等作为增殖及分化诱导因子 ,进行增殖培养、分化诱导 ;免疫组化法进行细胞性质鉴定。结果　加入bFGF、EGF增殖培养 72h可见细胞分裂相 (成纤维细胞样细胞 )。加入RA、BDNF、GDNF诱导 3d ,部分细胞有NSE、GFAP成分表达 ;第 10d可见有神经元、神经胶质形态样细胞形成 ,经细胞成分 (NSE、GFAP)鉴定证实为神经元、神经胶质细胞。结论　BMSC在体外培养条件下 ,经过bFGF、EGF、RA、BDNF、GDNF等因子的“程序性”作用 ,可以分化成神经元和神经胶质细胞

Bone marrow stromal cells differentiate into neuron and glial cells in vitro

Objective To explore the feasibility of differentiation into neural cells from bone marrow stromal cells(BMSC)in vitro,and to offer reference for the applying of BMSC.Methods BMSC were acquired from adult dog.Basic fibroblast growth factor(bFGF),epidermal growth factor(EGF),all trans retinoic acid(RA),brain derived neurotrophic factor (BDNF),glial cell line derived neurotrophic factor(GDNF)were served as proliferation and differentiation factors.Immunohistochemistry were used to identify the cells type.Results When bFGF and EGF had been added into the culture medium for 72 hour,cleavaged phase of larger flat fibroblast like cells apeared.On the 3rd day after adding RA,BDNF and GDNF into the culture medium,part of the BMSC derived cells started to express neuron specific enolase(NSE)or glial fibrillary acidic protein(GFAP).The neural cell modality like cells appeared on 10th day after adding inducing factors into the cultured medium,and that were proved by NSE or GFAP immunohisto chemical staining.Conclusion BMSC can differentiate into neural cells in vitro,induced programmatically by bFGF,EGF,RA,BDNF and GDNF.