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- Publisher
- Springer Journals
- Copyright
- Copyright © 2007 by Nature Publishing Group
- Subject
- Life Sciences; Life Sciences, general; Biological Techniques; Analytical Chemistry; Microarrays; Computational Biology/Bioinformatics; Organic Chemistry
- ISSN
- 1754-2189
- eISSN
- 1750-2799
- DOI
- 10.1038/nprot.2007.54
- Publisher site
- See Article on Publisher Site
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.
Journal
Nature Protocols – Springer Journals
Published: Mar 15, 2007