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大鼠胎儿神经干细胞的分离、培养及诱导分化
李 勇1, 沈文正1, 杨春荣1
西北农林科技大学 国家干细胞工程技术研究中心陕西分中心
摘要:
为了探讨神经干细胞(Neural stem cells,NSCs)在体外培养中的生物学特性,研究不同细胞因子对NSCs分化的影响,试验对大鼠胎儿神经干细胞进行了分离培养,并研究了10,50和100 ng/mL的神经生长因子(nerve growth factor,NGF)、脑源性神经营养因子(brain derived neruotrophic factor,BDNF)和胶质源性神经生长因子(glial cell line-derived neurotrophic factor,GDNF)对NSCs的定向诱导分化作用。结果表明,大鼠胎儿NSCs可在体外增殖并形成典型的神经球,指数增长期在传代后的第4~5天;10和50 ng/mL的NGF、BDNF和GDNF对NSCs的诱导分化作用不明显,100 ng/mL的NGF、BDNF和GDNF可诱导NSCs分化为神经元或胶质细胞。说明NGF和BDNF可诱导NSCs向神经元方向分化,GDNF诱导其向胶质细胞分化。
关键词:  大鼠  胎儿神经干细胞  神经生长因子  生长曲线
DOI:
分类号:
基金项目:国家“863”计划项目(2002AA216161)
Rat fetal neural stem cell separate, cultured and differentiated
Abstract:
To discuss the characteristics of neural stem cells biologic cultured in vitro and observe the effect of different growth factors on the differentiation of NSCs,the neural stem cells of rat fetal was seperated and cultured,and then they were directional induced by NGF,BDNF and GDNF in the concentration grads of 10 ng/mL,50 ng/mL and 100 ng/mL in the serum free medium and also identified.The result suggest that when cultured in vitro,NSCs can form typical spheres and proliferate,and the exponential growth occurred at the 4th-5th day after NSCs passaged,the effect of the neural factors was not obviously in low concentrations of 10 ng/mL or 50 ng/mL,but the NSCs can be differentiated into neurons and glial cells when they are cultured in the medium supplied with 100 ng/mL NGF,BDNF or GDNF.So in the serum free medium supplied with 20 ng/mL EGF and bFGF,the growth characteristic of NSCs would be definitely influenced,and NGF and BDNF could accelerate NSCs differentiating into neurons,however GDNF could induce them to glial cells.
Key words:  rat,NSCs,neurotrophic factors,growth curve