不同状态下新生鼠海马神经干细胞体外培养特征

目的探讨不同状态下新生鼠海马神经干细胞(NSCs)体外培养特征。方法将42只新生7日龄SD乳鼠随机分为正常对照组、单纯缺氧组及缺氧缺血(HI)组,每组14只。每组又根据处死时相点随机分成3、6 h,1、3、7、14、21 d 7个小组,每小组2只。建立HI动物模型后,采用细胞克隆和免疫组织化学技术对3组左侧海马的NSCs进行培养、传代、分化及鉴定。结果3组SD鼠海马组织均能培养出悬浮生长的NSCs神经球,具有连续克隆能力,可传代培养及分化。在3、6 h时间点处死培养SD鼠原代细胞时,3组获得克隆球的数目无显著差异,而在1、3、7、14、21 d时间点培养的原代细胞中均以单纯缺氧组培养出的克隆球最多,HI组培养出的克隆球最少。结论不同状态下新生鼠海马组织均能培养出NSCs,NSCs随年龄的增加、病程的延长和病情的加重而减少。

Characters of Culture of Neural Stem Cells from Hippocampus of Neonatal Rats in Different States in Vitro

Objective To investigate the characters of culture of neural stem cells(NSCs) from neonatal rats in different states in vitro.Methods Forty-two neonatal rats at age 7 days were divided randomly into 3 groups as control group,hypoxic group and hypoxic-ischemic group,each having 14 rats.Forteen rats of every group divided randomly into 7 small groups,each including 3 h,6 h,1 d,3 d,(7 d),14 d and 21 d,according to the time to put to death,each having 2 rats.After builting rat models of hypoxic-ischemic encephalopathy,NSCs from hippocampus in 3 groups were isolated,then cultured,passed,differentiated and differentiated with single cell clone and immunocytochemistry tecnique.Results NSCs in hippocampus from 3 groups were cultured in form of typical neuraospheres in suspension.The cells could be cloned,passed continuously and induced.There were differences among 3 groups when primary NSCs were cultured at 3 h and 6 h time points.But at 1 d,3 d,7 d,14 d and 21 d time points,clony neuraospheres from primary NSCs in hypoxic group were the most among 3 groups while clony neuraospheres from primary NSCs in hypoxic-ischemic group were the lest.Conclusions NSCs from hippocampus of neonatal rats in different states remain to be cultured,meanwhile,NSCs are decreased with increase of age,elongation of illness course and progress of state of an illness.