A protocol for isolation and culture of human umbilical vein endothelial cells
A protocol for isolation and culture of human umbilical vein endothelial cells
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DOI：
10.1038/nprot.2007.54
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发表时间：
2007-01-01
期刊：
影响因子：
14.8
通讯作者：
Vaubourdolle, Michel
中科院分区：
文献类型：
--
作者：
Baudin, Bruno;Bruneel, Arnaud;Vaubourdolle, Michel
我们描述了一种易于分离和培养人脐静脉内皮细胞（HUVECs）的方案，为每个研究人员提供了一种可以应用于细胞生物学实验室的方法。通过胶原酶处理从脐静脉血管壁分离内皮细胞（EC），然后接种在纤连蛋白包被的板上，并在37 ℃-5%CO2培养箱中在具有Earles盐和胎牛血清（FCS）但没有生长因子补充的培养基中培养7天。可以通过相差显微镜监测细胞融合;可以使用细胞表面或细胞内标记物表征EC并检查污染。各种方案可以应用于HUVEC，从简单的收获到蛋白质的特定溶解用于蛋白质组学分析。
We describe a protocol for easy isolation and culture of human umbilical vein endothelial cells (HUVECs) to supply every researcher with a method that can be applied in cell biology laboratories with minimum equipment. Endothelial cells (ECs) are isolated from umbilical vein vascular wall by a collagenase treatment, then seeded on fibronectin-coated plates and cultured in a medium with Earles' salts and fetal calf serum (FCS), but without growth factor supplementation, for 7 days in a 37 degrees C-5% CO2 incubator. Cell confluency can be monitored by phase-contrast microscopy; ECs can be characterized using cell surface or intracellular markers and checked for contamination. Various protocols can be applied to HUVECs, from simple harvesting to a particular solubilization of proteins for proteomic analysis.