title: A protocol for isolation and culture of human umbilical vein endothelial cells.
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 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.
authors.affiliation: 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. bruno.baudin@sat.aphp.fr
authors.forename: Bruno
authors.initials: B
authors.lastname: Baudin
authors.forename: Arnaud
authors.initials: A
authors.lastname: Bruneel
authors.forename: Nelly
authors.initials: N
authors.lastname: Bosselut
authors.forename: Michel
authors.initials: M
authors.lastname: Vaubourdolle
mesh_terms: D018929:Cell Culture Techniques / Q000379:methods*; D017364:Collagenases; D042783:Endothelial Cells / Q000166:cytology*; D006801:Humans; D008858:Microscopy, Phase-Contrast; D014471:Umbilical Veins / Q000166:cytology*
chemical_list: D017364:Collagenases