A protocol for isolation and culture of human umbilical vein endothelial cells

被引:368
|
作者
Baudin, Bruno [1 ,2 ]
Bruneel, Arnaud [2 ]
Bosselut, Nelly [2 ]
Vaubourdolle, Michel [2 ]
机构
[1] Univ Paris 11, UFR Pharm, UPRES JE 2493, Lab Biochim & Biol Cellulaire, F-92290 Chatenay Malabry, France
[2] Hop St Antoine, AP HP, F-75571 Paris 12, France
关键词
D O I
10.1038/nprot.2007.54
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
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.
引用
收藏
页码:481 / 485
页数:5
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