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目的探讨不同胎龄人胎脑室下区神经干细胞(NSCs)的生长特征。方法收集胎龄16～36周的胎儿90例,每例研究对象取室下区脑组织作为实验材料。所有研究对象产前经B超诊断患有先天性心脏病或消化道畸形,均无脑发育异常,孕妇及其丈夫要求终止妊娠。采用免疫组化技术对人胎脑室下区NSCs的形态、存在方式以及数量进行检测,采用细胞培养技术对其进行培养、传代、分化观察,并应用免疫组化技术对培养及分化的细胞进行鉴定。结果人胎脑室下区NSCs具有星形、圆形、椭圆形及梭形等形态,以星形最多。细胞胞浆相对较少;核呈圆形,染色质疏松,1～4个核仁不等。可见NSCs对称分裂和不对称分裂现象,NSCs多数单个散在分布,有的NSCs与别的NSCs形成突触联系。不同胎龄人胎脑室下区NSCs随着胎龄的增加而减少(X2=4644．602,P<0．01)。不同胎龄以及同一胎龄不同个体的NSCs在上述方面存在一定的差别。从人胎脑室下区分离的细胞在无血清培养时呈悬浮状态生长,形成神经球。该细胞可传代培养,表达神经巢蛋白抗原(Nestin);而在含血清培养时能分化,分化细胞表达神经元细胞、少突及星形胶质细胞的特异性抗原。结论不同胎龄人胎脑室下区均存在NSCs,且在形态、存在方式及数量上存在一定的差异。不同胎龄人胎脑室下区均能在体外培养出NSCs。
Objective To investigate the growth characteristics of neural stem cells (NSCs) in the subventricular zone of fetal human embryos of different gestational ages. Methods Totally 90 fetuses with gestational age ranging from 16 to 36 weeks were collected. The brain tissue of subventricular zone was taken as experimental material in each case. All subjects were diagnosed with prenatal B ultrasound with congenital heart disease or gastrointestinal malformations, no brain abnormalities, pregnant women and their husbands require termination of pregnancy. Immunohistochemistry was used to detect the shape, existence and number of NSCs in the subventricular zone of human fetus. The cells were cultured, passaged and differentiated by cell culture technique, and the cultured and differentiated cells were identified by immunohistochemistry . Results Human fetal subventricular zone of NSCs with stars, round, oval and fusiform and other forms, the most stars. Cytoplasm is relatively small; nucleus was round, chromatin loose, ranging from 1 to 4 nucleoli. NSCs can be seen symmetrical and asymmetric division of the phenomenon, the majority of NSCs scattered in a single, some NSCs and other NSCs synaptic connections. The NSCs in the subventricular zone in different gestational age human fetuses decreased with gestational age (X2 = 4644.602, P <0.01). There are some differences in the above aspects between NSCs of different gestational age and different gestational age. Cells isolated from the subependymal zone of the human fetus are suspended in a serum-free culture and form neurospheres. The cells can be passaged and cultured to express Nestin. When cultured in serum, the cells can differentiate and differentiate into neurons, oligodendrocytes and astrocytes. CONCLUSION NSCs are present in the subventricular zone of fetal human embryos at different gestational ages, and there are some differences in morphology, existence and quantity. Different fetal gestational age subventricular zone can be cultured in vitro NSCs.