Protocol abstract
Nature Protocols 2, - 481 - 485 (2007)
Published online: 15 March 2007 | doi:10.1038/nprot.2007.54
Subject Categories: Cell and tissue culture | Isolation, purification and separation
A protocol for isolation and culture of human umbilical vein endothelial cells
Bruno Baudin1,2, Arnaud Bruneel2, Nelly Bosselut2 & Michel Vaubourdolle2
Abstract
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 °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.
- Laboratoire de Biochimie et Biologie Cellulaire—UPRES JE 2493, UFR de Pharmacie, Université Paris-Sud 11, 3 rue Jean-Baptiste Clément, 92290 Châtenay-Malabry, France.
- Biochimie A, Hôpital Saint-Antoine, AP-HP, 184 rue du Faubourg Saint-Antoine, 75571, Paris Cedex 12, France.
Correspondence to: Bruno Baudin1,2 e-mail: bruno.baudin@sat.aphp.fr
