Yin Xiao-juan, Ju Rong, Feng Zhi-chun
Department of Pediatrics, Zhujiang Hospital, Nanfang Medical University, Guangzhou 510282, China.
Zhonghua Er Ke Za Zhi. 2006 Jul;44(7):500-4.
To study growth characteristics of neural stem cells (NSCs) from subventricular zone (SVZ) of the different human fetal brain at different gestational age and to provide experimental and theoretical evidences for clinical application of NSCs for treatment of certain diseases.
Ninety human embryos at gestational age 16 - 36 weeks were collected and were divided into six groups according to gestational age: 16 w, 20 w, 24 w, 28 w, 32 w and 36 w. Each group had 15 embryos and brain tissues were taken from each embryo's SVZ. All subjects had congenital heart disease or digestive tract abnormity diagnosed with B ultrasound at antepartum, but none had abnormal development of brain. Pregnant mother and her husband desire termination of pregnancy. The morphology, existing mode and the number of neural stem cells in subventricular zone were examined with immunohistochemical method. The NSCs in subventricular zone were cultured, passaged and differentiated with cell culture technique, then were identified with immunohistochemical method.
NSCs in SVZ from the different human fetal brain existed in a scattered manner in the network formed by stellate cells, NSCs had round, ellipse and fusiform shape, especially in stellate shape. NSCs had larger and smaller size and distributed in dense or scattered forms, each having zero to two enations, most had one or two. NSCs had less cytoplasm. The nucli of the NSCs had a round shape with loose chromatin and 1 - 4 nucleoli. Most of NSCs existed in singular scattered form, some of them showed symmetrical or asymmetrical division, some of them showed synaptic connection with other NSCs. The number of NSCs in SVZ from groups with different fetal age decreased with increasing gestational age (chi(2) = 4644.602, P < 0.01). NSCs in SVZ from the different human fetal brain cultured with serum-free medium formed typical neurospheres in suspension. The cells could be passaged continuously, and could express nestin antigen. Serum-contained medium induced neural stem cells to differentiate and express specific antigens of neuron, astrocyte and oligodendrocyte.
NSCs existed in SVZ of human embryos at different gestational age. There are differences in morphology, existing pattern and the number of NSCs in SVZ at different gestational age. NSCs in SVZ at different gestational age may be cultured in vitro.
研究不同胎龄人胎脑脑室下区（SVZ）神经干细胞（NSCs）的生长特性，为NSCs临床治疗某些疾病提供实验和理论依据。
收集90例孕16 - 36周的人胚胎，按胎龄分为6组：16周、20周、24周、28周、32周和36周。每组15例胚胎，取各胚胎SVZ脑组织。所有研究对象产前经B超诊断有先天性心脏病或消化道畸形，但无脑发育异常。孕妇及其丈夫要求终止妊娠。采用免疫组化方法检测脑室下区神经干细胞的形态、存在方式及数量。应用细胞培养技术对脑室下区NSCs进行培养、传代及分化，然后用免疫组化方法进行鉴定。
不同人胎脑SVZ的NSCs散在于星状细胞构成的网络中，呈圆形、椭圆形及梭形，以星状居多。NSCs大小不一，分布密集或稀疏，有0至2个突起，多数有1或2个突起。NSCs胞质较少。NSCs细胞核呈圆形，染色质疏松，有1 - 4个核仁。多数NSCs呈单个散在形式存在，部分表现为对称或不对称分裂，部分与其他NSCs有突触联系。不同胎龄组SVZ的NSCs数量随胎龄增加而减少（χ² = 4644.602，P < 0.01）。不同人胎脑SVZ的NSCs在无血清培养基中培养可形成典型的悬浮神经球。细胞可连续传代，并能表达巢蛋白抗原。含血清培养基诱导神经干细胞分化并表达神经元、星形胶质细胞及少突胶质细胞的特异性抗原。
不同胎龄人胚胎脑室下区存在NSCs。不同胎龄脑室下区NSCs在形态、存在形式及数量上存在差异。不同胎龄脑室下区NSCs可进行体外培养。
