神经干细胞植入鼠侧脑室后迁移与神经元分化  

Migration and Neuron Differentiation of Neural Stem Cells When Transplanted to the Lateral Ventricle of Neonatal Rat

作　　者：卜笑松[1] 栾佐[1] 尹国才[1] 唐久来[1] 

机构地区：[1]安徽医科大学第一附属医院儿科,合肥230022

出　　处：《实用儿科临床杂志》2007年第24期1852-1854,共3页Journal of Applied Clinical Pediatrics

摘　　要：目的探讨人胎脑神经干细胞(hNSC)在新生鼠(生后9d内)脑内迁移与分化,为hNSC移植治疗新生儿脑病提供实验依据。方法无血清培养基从流产人胎脑前脑培养出hNSC球,取部分同批hNSC球,用酶消化为单细胞悬液。将培养14d的hNSC和消化好的单细胞悬液经脑室途径移植到出生8d的健康SD大鼠侧脑室内,于移植24、48、72h通过取脑,制作石蜡切片,免疫组织化学观察。结果体外培养获得典型的人胎脑hNSC球,神经细胞呈球样悬浮生长,移植入脑室内的2种类型细胞,其在脑内存活迁移的结果明显不同,用抗人细胞核抗体(Anti-hNuc)免疫荧光检测,移植入细胞球的72h就可迁移到嗅球、额叶皮层的内侧中央前区、海马、枕叶皮层等部位的颗粒层内、小脑回内相当于蒲氏细胞层位置有排列整齐的单层细胞,中脑区域也有广泛的细胞分布;抗神经纤维抗体Anti-NF可看到大脑皮层的颗粒层和嗅球部有阳性反应细胞,细胞间通过突起发生了连接。移植入单细胞悬液组24h可在脑室内见到大量的Anti-hNuc(+)细胞,72h组可在脑室内及其附近脑组织内均见少量的细胞。结论体外培养获得的hNSC移植入鼠脑后可存活并发生迁移分化,细胞形态不同存活迁移分化的效果有明显不同,以直接培养出来的细胞球移植较制备成单细胞悬液移植的后存活迁移的能力明显增强。Objective To explore the migration and differentiation of the human neural stem cells （hNSC） after being transplanted to the neonatal rat lateral ventricle,to provide some data on therapy for neonatal cerebrepathy by using of neural stem cells. Methods N2 medium containing EGF ＋ FGF2 ＋ LIE was used to culture the NSC spheres from the forebrain tissues of aborted human fetus, The hNSC was identified by detecting the NSC marker nestin antigen and showing the potency to differentiate into neural cells（ including astrocytes,oligedendrocytes and neurons）by using indirect immuno-fluorescence assay（IFA）. The part of the hNSC in-vitro cultured for 14 d was digested to suspensions of cell. Cultured for 14 d, the hNSC in - vitro and the suspension were transplanted into the lateral ventricles of the neonatal rat brains. The rats were respect/rely killed at 24,48 and 72 h respectively post - transplant,the whole brain was sectiened,and the special immuno - response detection was performed by using anti -human nuclei（anti -hNuc）and anti -human neurofilament （anti - hNF）. Results In vitro culture,the typical NSC spheres were obtained from the forebrain of the human fetus. The suspensions of cells were obtained from the neurosphere. In neurosphere group, the results of anti - hNuc detecting tracing at 72 h post - injection showed that the grafts had migrated into the cortex grand layers of olfactory bulbns,mdial precentral area of lohus frentalis,hippocampal,and lobus oecipitalis. The label -positive cells lined along the Cerebellar Purkinje cell layers and appeared in most parts of mesencephalons. The immuro - respons resuhs of anti - hNF showed that the positive cells scattered in the grand layer of cortex,the connection among positive cells was watched. In suspensions group,the resuhs of anti - hNuc detecting tracing at 24 h post - injection show a great quantity of positive cells in the ventricles and injection track. At 72 h, a small quantity of positive cells remained in the ventricle and nearby b

关 键 词：人胎脑神经干细胞 移植 迁移 分化 

分 类 号：R329[医药卫生—人体解剖和组织胚胎学]