摘要
外培目的观察小鼠雪旺细胞体养时对大鼠神经干细胞存活及分化的影响。方法获取Wistar鼠坐骨神经并采用组织块法分离和纯化雪旺细胞；体外分离新生乳鼠脑神经干细胞，将雪旺细胞和神经干细胞分别培养扩增后进行共培养。共分5组进行：实验组1（NSC悬浮＋SC悬浮＋DMEM／F12）；实验组2（NSC悬浮＋SC贴壁＋DMEM／F12）；试验对照组1（SC培养基＋NSC＋DMEM／F12）；试验对照组2（EGF／bFGF＋NSC＋DMEM／F12）；试验对照组3（NSC＋DMEM／F12）。倒置相差显微镜对各组培养细胞每天观察形态和计数，免疫组织化学检测混合培养细胞特异性标记物的表达：SC采用P0和S100，NS采用nestin标记，神经干细胞分化神经元分别采用GFAP、GalC、Tubulin-β染色。结果共培养组NF染色阳性神经元样细胞的百分率明显高干其他各组；实验组1克隆球直径明显高于其他各组，其平均直径为8μm；实验组神经元样细胞突起的长度比对照组的长，3周长度差为26．5-67．3μm。结论大鼠神经干细胞与小鼠雪旺细胞共培养使两者不仅能够共生，而且雪旺细胞能显著促进体神经干细胞分化为神经元样细胞；神经干细胞分化神经元突起增长并且有序排列成轴索样结构。
Objective To evaluate the effect of mouse Schwann cells on the rat neural stem cells when they are co-cultured. Methods The sciatic nerve was obtained and the Schwann cells （SC） were isolated and purified. The neural stem cells （NSC） were obtained from the neonatal mouse brain. There were co-cultured. There were 5 groups： group Ⅰ （NSC suspension ＋ SC suspension ＋ DMEM/F12） ;group Ⅱ（NSC suspension ＋ SC adherence ＋ DMEM/F12） ; control group Ⅰ （SC medium ＋ NSC ＋ DMEM/ F12） ;control group Ⅱ （EGF/bFGF ＋ NSC ＋ DMEM/F12） ;control group Ⅲ （NSC ＋ DMEM/F12）. By using the immunohistochemical method, the expression of specific markers in the co-cultured cells was detected. The morphological changes of NSC were observed under a microscope. Results The majority of NSC protruded several elongate processes and the expression of tubullin-β was high, while only small amount of cells were positive for collagen. The positive rate of NF dyeing in group I was higher and the diameter of its clone was larger than other groups. Conclusion Schwann ceils can promote the survival, regeneration and differentiation of the neural stem cells in vitro.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2006年第1期81-83,共3页
Chinese Journal of Experimental Surgery
基金
国家自然科学基金资助项目(30271313)
天津市科委重点基金资助项目(0238021)
吴阶平医学基金共同资助项目
