摘要
目的:探讨脐血间充质干细胞(MSCs)向神经样细胞定向诱导分化的条件及方法,以明确其神经分化潜能。方法:将人脐血间充质干细胞体外培养扩增、保存后,取第5代的MSCs分别用维甲酸(RA)、维甲酸联合神经生长因子(NGF)诱导方法向神经样细胞诱导,诱导分化期间观察细胞形态变化,并比较表达神经元特异性烯醇化酶(NES)mRNA、胶质纤维酸性蛋白(GFAP)mRNA的差异。结果:两种不同诱导方法均能诱导细胞发生典型变化,对照组未发现神经样细胞生长。免疫组化法显示三种诱导方法诱导后均表达Nestin、NSE、GFAP,NGF+RA组多于RA组。Real-time RT-PCR显示诱导组及对照组均有NSE mRNA、GFAP mRNA表达,但两种诱导方法诱导后表达上调(P<0.05),添加NGF较单纯RA组上调明显(P<0.05)。结论:脐血MSCs经两种神经营养因子诱导法均可分化为神经样细胞,并表达神经细胞特异性标志物,其中添加NGF后较单纯应用RA诱导效果好。
Objective：To study the condition and methods of neuron-differentiation of ucmscs. Methods： UCMSCs were cultured and cryopreserved .UCMSCs of P5 were induced to differentiate into neural like cells using Retinoic Acid and Never Growth Factor. Morphological changes were monitored during the differentiating procedure. Real-time RT-PCR method was used to analyze NSE and GFAP mRNA expression of UCMSCs post differentiation between two methods.Results： In two group using differentiation medium, the typical neural-like cells can be found. The control group has none of this type cell growing. Immunohistochemistry showed that three different neurotrophic factors induced MSCs were positive for Nestin, NSE and GFAP. The number of positively stained cells was most in RA plus NGF group. Real-time RT-PCR results showed NES mRNA, GFAP mRNA expression could be seen in two groups and control group, while neural-induction can really increase the expression（P0.05）. The regulation of RA plus NGF group was better than RA group significantly（P0.05）. Conclussion： UCMSCs can differentiate into neuron like cells in vitro by different neurotrophic factor methods and express neural specific marker, indicating two capacity to differentiate into a neurophenotype and revealling two potential use for neural transplantation. Thereinto, the effect of RA plus NGF group was better than RA group significantly.
出处
《陕西医学杂志》
CAS
2010年第4期404-407,共4页
Shaanxi Medical Journal
