新生豚鼠海马神经干细胞分化为类毛细胞的体外实验

Induction of neural stem cells from newborn fetal guinea pig by fetal bovine serum and artificial perilymph

目的体外定向诱导新生豚鼠海马神经干细胞分化为内耳毛细胞。方法将原代培养的新生豚鼠海马来源的神经干细胞进行体外培养,分别置入含有10%胎牛血清和5%～15%人工外淋巴液培养基使其分化,并用免疫荧光化学,蛋白免疫印迹扫描电镜对分化的细胞进行鉴定。结果胎牛血清和人工外淋巴液均能诱导新生豚鼠海马神经干细胞分化为表达毛细胞特异抗体的细胞。结论神经干细胞在胎牛血清和人工外淋巴液中可以存活并分化出表达毛细胞特异抗体的细胞,为神经干细胞移植内耳治疗提供理论依据。

OBJECTIVE To observe the effects of fetal bovine serum （FBS） and artificial perilymph （APL） on the differentiation of neural stem cells from newborn fetal guinea pigs. METHODS FBS or APL were added to DMEM/F12-based medium, the growth and differentiation of NSCs were observed. After 3 weeks, the differentiations of NSCs were examined by immunofluorescence staining, Scanning electron microscope and Western Blotting. RESULTS 1 A total of 3.9 %～13.6 % of the cells treated with FBS orAPL culture medium expressed Myosin Ⅶa. 2 The differentiation of NSCs was verified by the positive results of hair cell marker Myosin Ⅶa through immunefluorescence staining, scanning electron microscope and Western Blot respectively. CONCLUSION Neural stem cells possess the potential to differentiate to hair cell phenotypes in both FBS and APL culture in vitro.