【摘要】：目的研究缺氧條件對(duì)骨髓來(lái)源內(nèi)皮祖細(xì)胞(BM-EPCs)生物學(xué)功能的影響。方法經(jīng)密度梯度離心法獲得大鼠骨髓來(lái)源單核細(xì)胞,接種在包被有大鼠玻連蛋白的培養(yǎng)皿中,加入內(nèi)皮細(xì)胞生長(zhǎng)培養(yǎng)基-2,于常規(guī)培養(yǎng)條件(37℃、5%CO2)下培養(yǎng),第4天時(shí),棄去未貼壁細(xì)胞,全量更換新鮮培養(yǎng)基,隨機(jī)分為常規(guī)培養(yǎng)組和缺氧培養(yǎng)組,常規(guī)培養(yǎng)組于常規(guī)培養(yǎng)條件下繼續(xù)培養(yǎng)至第7天;缺氧培養(yǎng)組分別于常規(guī)培養(yǎng)的第4、5、6天,于缺氧培養(yǎng)條件(1%O2+5%CO2+94%N2)下繼續(xù)培養(yǎng)72、48、24 h;細(xì)胞表面標(biāo)志物檢測(cè)并Dil標(biāo)記的乙酰化低密度脂蛋白和FITC標(biāo)記的荊豆凝集素1共同染色方法鑒定BM-EPCs,分別對(duì)各組培養(yǎng)至第7天的細(xì)胞進(jìn)行內(nèi)皮分化功能檢測(cè)(Matrigel As-say)和抗凋亡檢測(cè)(Annexin V/PI)。結(jié)果隨著缺氧處理時(shí)間延長(zhǎng),BM-EPCs早期凋亡率增加明顯。與常規(guī)培養(yǎng)組(0.89±0.20)%相比,缺氧培養(yǎng)24 h組早期凋亡率(1.33±0.07)%改變不明顯(P0.05),缺氧培養(yǎng)48 h組(3.25±0.12)%、72 h組(7.48±1.53)%,早期凋亡率明顯增加(P0.05)。與常規(guī)培養(yǎng)組比較,缺氧培養(yǎng)24 h組的體外血管形成能力增強(qiáng)(P0.01),缺氧培養(yǎng)48、72 h組體外血管形成能力明顯下降(P0.01)。結(jié)論缺氧預(yù)處理BM-EPCs24 h,BM-EPCs早期凋亡率增加不明顯,細(xì)胞體外血管形成能力明顯增強(qiáng)。
[Abstract]:Objective to study the effects of hypoxia on the biological function of bone marrow derived endothelial progenitor cells (BM-EPCs). Methods Mononuclear cells from rat bone marrow were obtained by density gradient centrifugation. The monocytes were inoculated in a culture dish coated with rat Glassin. The endothelial cell growth medium (-2) was added to the culture medium. The cells were cultured under conventional culture conditions (37 鈩，

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