ICR小鼠胚胎NSCs体外培养和诱导分化

In Vitro Culture and Induced Differentiation of Neural Stem Cells from ICR Fetal Mouse

利用表皮生长因子(EGF)和碱性成纤维生长因子(bFGF)的联合促进作用使原代胚胎皮层神经干细胞(Neural Stem Cells,NSCs)稳定增殖,以此来分离培养ICR小鼠胚胎NSCs,并在体外高效诱导成神经元。通过诱导分化的对照实验获得神经元分化比例最高的方法。免疫细胞化学不仅证实了培养的NSCs可表达Nestin并具有多分化潜能,还发现NSCs与支持细胞(SCs)共培养时神经元分化比例高达60%(P<5)。研究发现细胞生长因子EGF(20 ng/mL)和bFGF(20ng/mL)能有效促使NSCs在无血清条件下增殖,并且支持细胞促进其向神经元分化。

To isolate and culture the ICR fetal mouse neural stem cells（NSCs） and efficiently produce induced-neurons in vitro,this study utilised the synergistic effects of EGF and bFGF to promote the proliferation of primary fetal cortical NSCs.The approach with highest neuronal differentiation rate was confirmed through control induction experiments.Immunocytochemistry staining recorded that NSCs had the pluripotency,and the neuronal differentiation rate in pattern of co-culture NSCs and sertoli cells（SCs） was up to 60%（P〈5）.Therefore,it was proved that EGF（20 ng/mL） and bFGF（20 ng/mL） could efficiently boost the proliferation of NSCs in the serum-free medium,moreover,SCs contributed significantly to inducing NSCs into neurons.