人脑脊液对鼠神经干细胞增殖分化的影响

Effect of human cerebrospinal fluid on the proliferation and differentiation of rat neural stem cells

目的研究脑外伤血性脑脊液和脑积水清亮脑脊液对神经干细胞增殖分化的影响。方法从大鼠脑组织中分离培养神经干细胞,分别用人的两种脑脊液培养,分为试验组(血性脑脊液组)和对照组(脑积水清亮脑脊液组),于不同时间观察神经干细胞在两组脑脊液中的生长及增殖分化情况;并应用免疫细胞化学技术对两组脑脊液中神经干细胞的分化进行鉴定和比较。结果神经干细胞在两种脑脊液中均能存活并分化,部分细胞悬浮生长达十余天之久。加脑脊液培养第二天试验组NSE阳性细胞(神经元)的比例比对照组高,有统计学意义(P<0.0 5);培养第十天试验组GFAP阳性细胞(星形胶质细胞)的比例比对照组高,NSE阳性细胞的比例比对照组低,有统计学意义(P<0.0 5)。结论神经干细胞在脑外伤血性脑脊液和脑积水清亮脑脊液中均能存活、增殖和分化。两组脑脊液对神经干细胞分化的种类上有差异。

Objective To investigate the effect of the injured brain cerebrospinal fluid（CSF） from traumatic brain injury and the hydrocephalus patients on the survival,proliferation and differentiation of neural stem cells（NSCs）.Methods NSCs were isolated and cultured from brain of SD rat.The NSCs were cultured in human bloody CSF from the injured brain patients（the experimental group） or CSF from the hydrocephalus patients（the control group）.The growth,proliferation and differentiation pattern of NSCs in two groups were observed at different times.The differentiated cells were identified by immunocytochemistry and their proportions were compared.Results The NSCs could survive and differentiate both in bloody CSF and hydrocephalus CSF,and part of the cells could grow up by floating in CSF more than ten days.On the second day of the cultivate process,the percentage of NSE positive cells（neurons） in experimental group is significantly higher than that of control group（P0.05）.On the tenth day,the percentage of GFAP positive cells（astrocytes） in the experimental group is significantly higher than that of control group,while the percentage of NSE positive cells in the experimental group is significantly lower than that of control group（P0.05）.Conclusions The NSCs can survive,proliferate and differentiate both in the bloody CSF and hydrocephalus CSF of injured brain.There was different effect on the differentiation of NSCs between the two groups.