Abstract
This protocol describes a simple and an economical method for isolation of endothelial cells from human umbilical vein. Umbilical cord is easily available postpartum following informed consent, and the method for its collection is noninvasive with few ethical concerns. Thus, umbilical vein is an ideal source for isolation of endothelial cells of human origin. Endothelial cells are isolated from umbilical vein by collagenase digestion. This is followed by their culture on extracellular matrix (ECM) protein coated tissue culture flasks in presence of endothelial cell growth medium. In the last few decades, human umbilical vein endothelial cells (HUVECs) have come to be regarded as a standard model system by vascular biologists to understand general principles of endothelial cell biology and dysfunction. Here, we describe isolation, culture, as well as validation of HUVECs.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Similar content being viewed by others
References
Jaffe EA (1987) Cell biology of endothelial cells. Hum Pathol 18(3):234–239
Michiels C (2003) Endothelial cell functions. J Cell Physiol 196(3):430–443
Pober JS, Sessa WC (2007) Evolving functions of endothelial cells in inflammation. Nat Rev Immunol 7(10):803–815
Sandoo A, van Zanten JJ, Metsios GS, Carroll D, Kitas GD (2010) The endothelium and its role in regulating vascular tone. Open Cardiovasc Med J 4:302–312
Davignon J, Ganz P (2004) Role of endothelial dysfunction in atherosclerosis. Circulation 109(23 Suppl 1):III27–III32
Rajendran P, Rengarajan T, Thangavel J, Nishigaki Y, Sakthisekaran D, Sethi G, Nishigaki I (2013) The vascular endothelium and human diseases. Int J Biol Sci 9(10):1057–1069
Liao JK (2013) Linking endothelial dysfunction with endothelial cell activation. J Clin Invest 123(2):540–541
Michiels C, Arnould T, Remacle J (2000) Endothelial cell responses to hypoxia: initiation of a cascade of cellular interactions. Biochim Biophys Acta 1497(1):1–10
Schwartz SM (1978) Selection and characterization of bovine aortic endothelial cells. In Vitro 14(12):966–980
Fehrenbach ML, Cao G, Williams JT, Finklestein JM, Delisser HM (2009) Isolation of murine lung endothelial cells. Am J Physiol Lung Cell Mol Physiol 296(6):L1096–L1103
Ruck T, Bittner S, Epping L, Herrmann AM, Meuth SG (2014) Isolation of primary murine brain microvascular endothelial cells. J Vis Exp no. 93:e52204
Jaffe EA, Nachman RL, Becker CG, Minick CR (1973) Culture of human endothelial cells derived from umbilical veins. Identification by morphologic and immunologic criteria. J Clin Invest 52(11):2745–2756
Baudin B, Bruneel A, Bosselut N, Vaubourdolle M (2007) A protocol for isolation and culture of human umbilical vein endothelial cells. Nat Protoc 2(3):481–485
Cheung AL (2007) Isolation and culture of human umbilical vein endothelial cells (HUVEC). Curr Protoc Microbiol Appendix 4:Appendix 4B
Onat D, Brillon D, Colombo PC, Schmidt AM (2011) Human vascular endothelial cells: a model system for studying vascular inflammation in diabetes and atherosclerosis. Curr Diab Rep 11(3):193–202
Bachetti T, Morbidelli L (2000) Endothelial cells in culture: a model for studying vascular functions. Pharmacol Res 42(1):9–19
Bishop ET, Bell GT, Bloor S, Broom IJ, Hendry NF, Wheatley DN (1999) An in vitro model of angiogenesis: basic features. Angiogenesis 3(4):335–344
Giri H, Chandel S, Dwarakanath LS, Sreekumar S, Dixit M (2013) Increased endothelial inflammation, sTie-2 and arginase activity in umbilical cords obtained from gestational diabetic mothers. PLoS One 8(12):e84546
Pantham P, Aye IL, Powell TL (2015) Inflammation in maternal obesity and gestational diabetes mellitus. Placenta 36(7):709–715
Redman CW, Sacks GP, Sargent IL (1999) Preeclampsia: an excessive maternal inflammatory response to pregnancy. Am J Obstet Gynecol 180(2 Pt 1):499–506
Jaffe EA, Hoyer LW, Nachman RL (1974) Synthesis of von Willebrand factor by cultured human endothelial cells. Proc Natl Acad Sci USA 71(5):1906–1909
Larrivee B, Karsan A (2005) Isolation and culture of primary endothelial cells. Methods Mol Biol 290:315–329
Garlanda C, Dejana E (1997) Heterogeneity of endothelial cells. Specific markers. Arterioscler Thromb Vasc Biol 17(7):1193–1202
Voyta JC, Via DP, Butterfield CE, Zetter BR (1984) Identification and isolation of endothelial cells based on their increased uptake of acetylated-low density lipoprotein. J Cell Biol 99(6):2034–2040
Adachi H, Tsujimoto M, Arai H, Inoue K (1997) Expression cloning of a novel scavenger receptor from human endothelial cells. J Biol Chem 272(50):31217–31220
Szulman AE (1962) The histological distribution of the blood group substances in man as disclosed by immunofluorescence: II. The H antigen and its relation to A and B antigens. J Exp Med 115(5):977–996
Szulman AE (1964) The histological distribution of the bloop group substances in man as disclosed by immunofluorescence: III. The A, B, and H antigens in embryos and fetuses from 18 mm in length. J Exp Med 119:503–516
Szulman AE (1972) The A, B and H blood-group antigens in human placenta. N Engl J Med 286(19):1028–1031
Cantelmo AR, Brajic A, Carmeliet P (2015) Endothelial metabolism driving angiogenesis: emerging concepts and principles. Cancer J 21(4):244–249
Lamalice L, Le BF, Huot J (2007) Endothelial cell migration during angiogenesis. Circ Res 100(6):782–794
Goldsmith JC, McCormick JJ, Yen A (1984) Endothelial cell cycle kinetics. Changes in culture and correlation with endothelial properties. Lab Investig 51(6):643–647
Sow RC (2012) Introduction to Cell culture: Culture of human endothelial cells from umbilical veins. Methods Mol Biol 806: 265-274.
Acknowledgments
The authors acknowledge funding received from the Department of Biotechnology (DBT) [File# BT/PR12547/MED/30/1456/2014] and Science and Engineering Research Board (SERB) [File# EMR/2015/000704], Government of India for their work.
Author information
Authors and Affiliations
Corresponding author
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2024 Springer Science+Business Media, LLC, part of Springer Nature
About this protocol
Cite this protocol
Chandel, S., Kumaragurubaran, R., Giri, H., Dixit, M. (2024). Isolation and Culture of Human Umbilical Vein Endothelial Cells (HUVECs). In: Tharakan, B. (eds) Vascular Hyperpermeability. Methods in Molecular Biology, vol 2711. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3429-5_12
Download citation
DOI: https://doi.org/10.1007/978-1-0716-3429-5_12
Published:
Publisher Name: Humana, New York, NY
Print ISBN: 978-1-0716-3428-8
Online ISBN: 978-1-0716-3429-5
eBook Packages: Springer Protocols