【摘要】：目的建立一個詳細(xì)、易于重復(fù)的人臍靜脈內(nèi)皮細(xì)胞原代培養(yǎng)操作流程。方法采用膠原酶消化法分離人臍靜脈內(nèi)皮細(xì)胞;用倒置顯微鏡觀察細(xì)胞形態(tài),免疫熒光法檢測內(nèi)皮細(xì)胞標(biāo)志分子,Western blotting檢測內(nèi)皮細(xì)胞標(biāo)志酶——一氧化氮合酶的含量。結(jié)果 0.2%膠原酶Ⅰ在37℃條件下消化5~13 min可得到高純度(95%)人臍靜脈內(nèi)皮細(xì)胞;在倒置顯微鏡下,人臍靜脈內(nèi)皮細(xì)胞呈典型的鵝卵石樣,細(xì)胞核明顯;Ⅷ因子和CD31的免疫熒光檢測呈陽性;Western blotting結(jié)果顯示,分離到的人臍靜脈內(nèi)皮細(xì)胞中有一氧化氮合酶持續(xù)合成。結(jié)論建立的人臍靜脈內(nèi)皮細(xì)胞原代培養(yǎng)和鑒定的操作流程簡單易學(xué),且在形態(tài)、標(biāo)志分子和關(guān)鍵性標(biāo)志酶3個方面提供細(xì)胞鑒定的方法和參考數(shù)據(jù)。
[Abstract]:Objective to establish a detailed and reproducible primary culture procedure for human umbilical vein endothelial cells (HUVECs). Methods Human umbilical vein endothelial cells were isolated by collagenase digestion, the morphology of endothelial cells was observed by inverted microscope, and the content of nitric oxide synthase (NOS), a marker of endothelial cells, was detected by immunofluorescence assay. Results 0.2% collagenase I could obtain high purity (95%) of human umbilical vein endothelial cells after digesting at 37 鈩，

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