中文摘要：

目的探讨缺氧对体外培养的大鼠神经干细胞（neuralstemceils，Nscs）增殖、分化和细胞凋亡的影响。方法分离成体SD大鼠室管膜室下区（subventricularzone，sVz）组织，NSCs在常氧和低氧培养箱培养，光镜观察细胞形态；采用NSCs标记物（nestin）免疫荧光染色鉴定细胞，未成熟神经元标记物（β-tubulinⅢ/Tju-1）、星形胶质细胞标记物（GFAP）和少突胶质细胞标记物（04）免疫荧光染色和细胞计数观察细胞的分化能力；WST-1检测细胞增殖能力；流式细胞仪和TUNEL法检测细胞凋亡。结果从成体SVZ组织中成功分离培养出NSCs；无血清诱导下细胞培养至第4代有部分细胞分化，多为胶质细胞；血清诱导后NSCs可以向神经元、星形胶质细胞和少突胶质细胞分化，大部分干细胞分化为TjU一1阳性细胞（53．8％±6．8％），部分为GFAP阳性细胞（21．6％±5．4％），少部分为04阳性细胞（11．4％±4．5％）。一定时间（〈24h）的低氧（3mL／L氧浓度）可以促进NSCs增殖（P〈0．05），但长时间缺氧（〉24h），细胞增殖能力减弱而细胞凋亡比例增加（P〈O．01）。结论正常成体组织中存在NSCs；体外培养条件下分离培养的NSCs可以分化为多种类型细胞；短时间低氧可促进NSCs的增殖，而长时间缺氧导致细胞凋亡比例增加。

英文摘要：

Objective To explore the effect of hypoxia on the proliferation, differentiation and apoptosis of neural stem cells （NSCs） isolated from adult rat subventricular zone （SVZ） and cultured in vitro. Methods The NSCs isolated from adult rat SVZ were cultured under normoxia condition or hypoxia condition （hypoxia for 6, 12, 24, 48 or 72 h） in vitro. The nestin＋ cells, Tju-1＋ cells, GFAP＋ cells and 04＋ cells were detected by immunofluorescence staining, and the percentage of the positive cells was measured respectively. The proliferation of the NSCs was assayed with cell proliferation reagent （WST-1） and the cell cycle was detected by flow cytometry （FCM）. The apoptosis of the NSCs was detected by TUNEL and flow cytometry. Results The NSCs from adult rat SVZ were successfully isolated and cultured in vitro. Induced by serum, the NSCs differentiated at the 4th generation and most of them were Tju-l-positive （53.8± 6.8%） while some of them were GFAP-positive （21.6 ± 5.4 %）. Besides, a small number of them were O4-positive （11.4± 4.5 % ）. Hypoxia （0.3 % 02 ） for a short period （~24 h） promoted the proliferation of the NSCs （P~ 0. 05）. But prolonged hypoxia （~ 24 h） reduced the proliferation capacity of the NSCs and increased the percentage of apoptotic cells （P〈0.01）. Gonclusion NSCs are present in normal adult tissues; NSCs isolated and cultured in vitro can differentiate into various types of cells. Short-term hypoxia can promote the proliferation of NSCs, but prolonged hypoxia induces apoptosis of NSCs.