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Vascular Cell

, 6:7

First Online: 01 April 2014Received: 30 December 2013Accepted: 10 March 2014

Abstract

BackgroundThe understanding of endothelial cell biology has been facilitated by the availability of primary endothelial cell cultures from a variety of sites and species; however, the isolation and maintenance of primary mouse aortic endothelial cells MAECs remain a formidable challenge.
Culturing MAECs is difficult as they are prone to phenotypic drift during culture.
Therefore, there is a need to have a dependable in vitro culture system, wherein the primary endothelial cells retain their properties and phenotypes.

MethodsHere, we developed an effective method to prepare immortalized MAEC iMAEC lines.
Primary MAECs, initially isolated from aortic explants, were immortalized using a retrovirus expressing polyoma middle T-antigen.
Immortalized cells were then incubated with DiI-acetylated-low density lipoprotein and sorted via flow cytometry to isolate iMAECs.

ResultsiMAECs expressed common markers of endothelial cells, including PECAM1, eNOS, VE-cadherin, and von Willebrand Factor.
iMAECs aligned in the direction of imposed laminar shear and retained the ability to form tubes.
Using this method, we have generated iMAEC lines from wild-type and various genetically modified mice such as p47, eNOS, and caveolin-1.

ConclusionIn summary, generation of iMAEC lines from various genetically modified mouse lines provides an invaluable tool to study vascular biology and pathophysiology.

KeywordsMAEC Endothelial cells Shear stress p47phox eNOS cav1 Electronic supplementary materialThe online version of this article doi:10.1186-2045-824X-6-7 contains supplementary material, which is available to authorized users.

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Author: Chih-Wen Ni - Sandeep Kumar - Casey J Ankeny - Hanjoong Jo

Source: https://link.springer.com/article/10.1186/2045-824X-6-7



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