A new, rapid and reproducible method to obtain high quality endothelium in vitro.

Cytotechnology
Nuria JiménezJan A Post

Abstract

Human umbilical vein endothelial cells (HUVECs) cultured in vitro are a commonly used experimental system. When properly differentiated they acquire the so-called cobblestone phenotype; thereby mimicking an endothelium in vivo that can be used to shed light on multiple endothelial-related processes. In the present paper we report a simple, flexible, fast and reproducible method for an efficient isolation of viable HUVECs. The isolation is performed by sequential short trypsinization steps at room temperature. As umbilical cords are often damaged during labor, it is noteworthy that this new method can be applied even to short pieces of cord with success. In addition, we describe how to culture HUVECs as valid cobblestone cells in vitro on different types of extracellular matrix (basement membrane matrix, fibronectin and gelatin). We also show how to recognize mature cobblestone HUVECs by ordinary phase contrast microscopy. Our HUVEC model is validated as a system that retains important features inherent to the human umbilical vein endothelium in vivo. Phase contrast microscopy, immuno-fluorescence and electron microscopy reveal a tight cobblestone monolayer. Therein cells show Weibel-Palade bodies, caveolae and junctional comple...Continue Reading

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Citations

Feb 25, 2014·Analytical Biochemistry·Michael B LoMonaco, Charles J Lowenstein
Jan 20, 2017·Biomedical Materials·Pierre-Antoine FayeAlexis Desmoulière
Jan 10, 2020·BioMed Research International·Hao DingYuling Gu
Dec 13, 2016·Advanced Functional Materials·Kathy Ye MorganLisa E Freed
Dec 6, 2018·International Journal of Molecular Sciences·Effat AlizadehHelder André
Feb 23, 2021·Frontiers in Bioengineering and Biotechnology·Giulia SilvaniBoris Martinac
Jun 9, 2021·Nature Communications·Chih-Yu WuHsien-Yeh Chen
Jul 3, 2021·Micromachines·Giulia GrisantiCarlo Massimo Casciola
Sep 26, 2019·Biomacromolecules·Antonino PuglisiYusuf Yagci

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Methods Mentioned

BETA
electron microscopy
phase contrast microscopy
scanning electron microscopy
gel filtration
protein determination
transmission

Software Mentioned

LAS EZ

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