本文選題：缺氧 + 室管膜室下區(qū)　； 參考：《西安交通大學(xué)學(xué)報(bào)(醫(yī)學(xué)版)》2013年06期

【摘要】：目的探討缺氧對(duì)體外培養(yǎng)的大鼠神經(jīng)干細(xì)胞(neural stem cells,NSCs)增殖、分化和細(xì)胞凋亡的影響。方法分離成體SD大鼠室管膜室下區(qū)(subventricular zone,SVZ)組織,NSCs在常氧和低氧培養(yǎng)箱培養(yǎng),光鏡觀察細(xì)胞形態(tài);采用NSCs標(biāo)記物(nestin)免疫熒光染色鑒定細(xì)胞,未成熟神經(jīng)元標(biāo)記物(β-tubulinⅢ/Tju-1)、星形膠質(zhì)細(xì)胞標(biāo)記物(GFAP)和少突膠質(zhì)細(xì)胞標(biāo)記物(O4)免疫熒光染色和細(xì)胞計(jì)數(shù)觀察細(xì)胞的分化能力;WST-1檢測(cè)細(xì)胞增殖能力;流式細(xì)胞儀和TUNEL法檢測(cè)細(xì)胞凋亡。結(jié)果從成體SVZ組織中成功分離培養(yǎng)出NSCs;無(wú)血清誘導(dǎo)下細(xì)胞培養(yǎng)至第4代有部分細(xì)胞分化,多為膠質(zhì)細(xì)胞;血清誘導(dǎo)后NSCs可以向神經(jīng)元、星形膠質(zhì)細(xì)胞和少突膠質(zhì)細(xì)胞分化,大部分干細(xì)胞分化為Tju-1陽(yáng)性細(xì)胞(53.8%±6.8%),部分為GFAP陽(yáng)性細(xì)胞(21.6%±5.4%),少部分為O4陽(yáng)性細(xì)胞(11.4%±4.5%)。一定時(shí)間(24h)的低氧(3mL/L氧濃度)可以促進(jìn)NSCs增殖(P0.05),但長(zhǎng)時(shí)間缺氧(24h),細(xì)胞增殖能力減弱而細(xì)胞凋亡比例增加(P0.01)。結(jié)論正常成體組織中存在NSCs;體外培養(yǎng)條件下分離培養(yǎng)的NSCs可以分化為多種類型細(xì)胞;短時(shí)間低氧可促進(jìn)NSCs的增殖,而長(zhǎng)時(shí)間缺氧導(dǎo)致細(xì)胞凋亡比例增加。
[Abstract]:Objective to investigate the effects of hypoxia on proliferation, differentiation and apoptosis of neural stem cells in vitro.Methods cultured adult SD rats subventricular zonetin (SVZ) tissues were cultured in normoxic and hypoxic incubators, the morphology of cells was observed under light microscope, and the cells were identified by immunofluorescence staining with NSCs labeling.Immature neuronal markers (尾 -tubulin 鈪，

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