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Histochem Cell Biol. 2019 Nov;152(5):377-390. doi: 10.1007/s00418-019-01815-2. Epub 2019 Sep 21.

Outgrowth, proliferation, viability, angiogenesis and phenotype of primary human endothelial cells in different purchasable endothelial culture media: feed wisely.

Author information

1
Department of Obstetrics and Gynaecology, Medical University of Graz, Graz, Austria.
2
Institute of Pathology, Medical University of Graz, Graz, Austria.
3
Omics Center Graz, BioTechMed-Graz, Graz, Austria.
4
Institute of Chemical Technologies and Analytics, Vienna University of Technology, Vienna, Austria.
5
Division of Bioinformatics, Biocenter, Medical University of Innsbruck, Innsbruck, Austria.
6
Department of Obstetrics and Gynaecology, Medical University of Graz, Graz, Austria. ursula.hiden@medunigraz.at.

Abstract

Function and dysfunction of endothelial cells are regulated by a multitude of factors. Endothelial cell research often requires in vitro cell culture experiments. Hence, various culture media specifically designed to promote endothelial cell growth are available. These strikingly differ in their composition: complex media contain endothelial cell growth supplement (ECGS), an extract produced of bovine brain with undefined amounts of biologically active compounds, whilst defined media contain selected growth factors in defined concentrations. We here compared the effect of seven purchasable endothelial cell culture media on colony outgrowth, proliferation, viability, in vitro angiogenesis and phenotype of mature primary human endothelial cells using feto-placental endothelial cells isolated from chorionic arteries (fpEC). The effect of media on colony outgrowth was additionally tested on umbilical cord blood-derived endothelial progenitor cells (ECFCs). Outgrowth, purity, proliferation and viability differed between media. Outgrowth of fpEC and ECFCs was best in a defined medium containing EGF, FGF2 and VEGF. By contrast, established fpEC isolations proliferated best in complex media containing ECGS, heparin and ascorbic acid. Also viability of cells was higher in complex media. In vitro angiogenesis was most intense in a defined medium containing the highest number of individual growth factors. FACS analysis of surface markers for endothelial cell subtypes revealed that endothelial phenotype of fpEC was unaffected by media composition. Our data demonstrate the fundamental effect of endothelial cell culture media on primary cell isolation success and behaviour. Whether the composition of supplements is suitable also for individual experiments needs to be tested specifically.

KEYWORDS:

Angiogenesis; Culture media; Outgrowth; Primary endothelial cells; Proliferation; Viability

PMID:
31541300
PMCID:
PMC6842357
DOI:
10.1007/s00418-019-01815-2
[Indexed for MEDLINE]
Free PMC Article

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