摘要
为探讨缺氧对体外培养的神经干细胞生长、分化及凋亡的影响,本研究采用显微解剖、机械吹打、无血清悬浮培养方法分离培养神经干细胞,用巢蛋白(nestin)免疫荧光染色对其进行鉴定。三气培养箱予以缺氧干预,分为5%缺氧组、10%缺氧组和正常对照组,每组又依缺氧干预时间的不同,分为6、12、24、48、72、96和120h组。通地绘制细胞生长曲线(MTT法)和计数克降形成率检测缺氧对神经干细胞增殖的影响。缺氧培养后,再用含10%胎牛血清的培养基进行分化培养,用免疫荧光技术检测缺氧对神经干细胞分化、凋亡及形态变化的影响。结果显示:缺氧干预后神经干细胞的增殖率明显下降,细包凋亡数增加,分化的神经元和胶质细胞的突起变短变粗,数量减少,但神经元和胶质细胞的分化比例未见明显变化。结果提示:缺氧可严重影响神经干细胞的存活和正常分化、且影响程序与缺氧剂量有量效关系。
This study was designed to explore the effects of hypoxia on proliferation, differentiation and apoptosis of neural stem cells （NSCs） from neural tube of embryonic rats. The NSCs were isolated by microdissection and mechanical blowing, cultured with serum-free suspension culture, and identified by immunofluoreseent staining using antibodies against nestin, NSE and GFAP. NSCs were exposed to 5% and 10% 02 for6 h, 12 h, 24 h, 48 h, 72 h, 96 h, or 120 h. Proliferation of NSCs were observed after hypoxia by MTT method and calculating clones. After NSCs were exposed to hypoxia with 10% fetal bovine serum（FBS） , the ratio of the differentiated neurons to glia cells, cell apoptosis and morphological changes were detected. The results showed that proliferation of NSCs decreased significantly after hypoxia, while the number of apoptotic cells increased. The differentiated neurons and glia decreased in number and their neurites became shorter and thinner. There were no significant changes of the ratio of the differentiated neurons to gila cells. The present results suggest that hypoxia may have great effect on the survival and differeatiation of NSCs in a dose-dependent manner.
出处
《神经解剖学杂志》
CAS
CSCD
北大核心
2006年第1期50-54,共5页
Chinese Journal of Neuroanatomy
关键词
神经干细胞
缺氧
增殖
分化
凋亡
neural stem cells, hypoxia, proliferation, differentiation, apoptosis
