人脐静脉内皮细胞体外培养、鉴定与形态学特征观察

Culture,identification and morphologic observation of endothelial cells from human unbilical vein in vitro

目的:建立人血管内皮细胞的培养模型,为体外研究动脉硬化发病机制提供重要的实验手段。方法:采用1.25g/L胰蛋白酶(2.5g/L胰蛋白酶:0.2g/LEDTA=1∶1V/V)消化、分离和传代人脐静脉内皮细胞,并用光镜、电镜和免疫组化等方法进行内皮细胞的形态观察和鉴定。结果:原代培养的内皮细胞在接种后约2h开始贴壁生长,光镜下内皮细胞呈铺路石状镶嵌排列;免疫组化可见内皮细胞胞浆中人第VIII因子相关抗原呈阳性反应;电镜下可见胞浆中有W-P小体,证实培养的细胞为内皮细胞。结论:用胰酶灌注消化脐静脉可获得高纯度的内皮细胞,细胞存活率高。本方法为血管内皮细胞的研究提供了实验模型。

AIM:To establish the model of human vascular endothelial cells(ECs) to provide important experimental method for the pathogenesis research of atherosclerosis in vitro. METHODS:ECs in umbilical vein were digested separated and subcultured with 1.25 g/L trypsin,(2.5 g/L trypsin:0.2 g/L EDTA=1:1V/V).And morphologic observation and evaluation of ECs were made with light microscopy,electron microscopy and immunohistochemistry. RESULTS:The primary cultured ECs started to grow attaching to flask about 2 hours after inoculating.The ECs arrayed like pitching stone under light microscopy;Ⅷfactor in the ECs showed positive reaction by immunohistochemistry;W P corpuscles was in the cell plasm under electron microscopy,which were identified as ECs. CONCLUSION:High purity EC can be harvested with digestion of pancreatin perfusion with high survival rate.This method provides experimental model for vascular EC research.