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

Nature Protocols volume 2, pages 481–485 (2007)Cite this article

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.

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Authors and Affiliations

  1. Laboratoire de Biochimie et Biologie Cellulaire—UPRES JE 2493, UFR de Pharmacie, Université Paris-Sud 11, 3 rue Jean-Baptiste Clément, Châtenay-Malabry, 92290, France

    Bruno Baudin

  2. Biochimie A, Hôpital Saint-Antoine, AP-HP, 184 rue du Faubourg Saint-Antoine, Paris, 75571, Cedex 12, France

    Bruno Baudin, Arnaud Bruneel, Nelly Bosselut & Michel Vaubourdolle

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  4. Michel Vaubourdolle
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Correspondence to Bruno Baudin.

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Baudin, B., Bruneel, A., Bosselut, N. et al. A protocol for isolation and culture of human umbilical vein endothelial cells. Nat Protoc 2, 481–485 (2007). https://doi.org/10.1038/nprot.2007.54

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