Department of Orthopaedics, Renmin Hospital, Wuhan University, No. 238, Jiefang Road, Wuchang District, Wuhan 430060, Hubei, China.
Neurol Sci. 2013 May;34(5):735-9. doi: 10.1007/s10072-012-1124-8. Epub 2012 Jun 5.
The purpose was to study the effect of human cerebrospinal fluid (CSF) on differentiation of rat neural stem cells (NSCs), and thus explore the feasibility of transplanting stem cells via lumbar puncture clinically. Rat NSCs derived from fetal brain were divided into two groups, and cultured in DMEM/F12 supplemented with 10 % FBS and human CSF, respectively. Cellular growth was observed with an inverted microscope, and immunostaining was used to analyze differentiation of NSCs in both groups. Cells of fetal brain showed shapes of spindle or star with minor sprouts at fifth day post-culture, and stained with nestin. NSCs in the control group differentiated into neurons, with positive staining to NSE, when cultured further in DMEM/F12 supplemented with 10 % FBS. While NSCs in the experiment group, cultured in CSF, differentiated into astroglia on eighth day, with positive immunostaining to GFAP. The new neurons dissolved rapidly when they were cultured in CSF. Human CSF cannot promote NSCs to differentiation toward neuron, nor support newborn neurons survival. It seems an inappropriate approach to transplant stem cells through CSF.
目的是研究人脑脊液(CSF)对大鼠神经干细胞(NSC)分化的影响,从而探讨经腰椎穿刺临床移植干细胞的可行性。将来源于胎鼠脑的 NSCs 分为两组,分别在添加 10%FBS 的 DMEM/F12 和人 CSF 中培养。倒置显微镜下观察细胞生长情况,并用免疫组化法分析两组 NSCs 的分化情况。培养第 5 天,胎脑细胞呈梭形或星形,有少量突起,巢蛋白染色阳性。继续在添加 10%FBS 的 DMEM/F12 中培养时,对照组 NSCs 分化为神经元,NSE 染色阳性。而实验组 NSCs 在 CSF 中培养 8 天后分化为星形胶质细胞,GFAP 免疫组化染色阳性。在 CSF 中培养时,新生神经元溶解迅速。人 CSF 不能促进 NSCs 向神经元分化,也不能支持新生神经元存活。通过 CSF 移植干细胞似乎不是一种合适的方法。