Liver Sinusoidal Endothelial Cells Escape Senescence by Loss of p19ARF.
Liver sinusoidal endothelial cells (LSECs) represent a highly differentiated cell type that lines hepatic sinusoids. LSECs form a discontinuous endothelium due to fenestrations under physiological conditions, which are reduced upon chronic liver injury. Cultivation of rodent LSECs associates with a...
Main Authors: | , , |
---|---|
Format: | Article |
Language: | English |
Published: |
Public Library of Science (PLoS)
2015-01-01
|
Series: | PLoS ONE |
Online Access: | http://europepmc.org/articles/PMC4631446?pdf=render |
id |
doaj-712729bc2a4c4ab7a9febd4fadf9af30 |
---|---|
record_format |
Article |
spelling |
doaj-712729bc2a4c4ab7a9febd4fadf9af302020-11-25T00:20:23ZengPublic Library of Science (PLoS)PLoS ONE1932-62032015-01-011011e014213410.1371/journal.pone.0142134Liver Sinusoidal Endothelial Cells Escape Senescence by Loss of p19ARF.Petra KoudelkovaGerhard WeberWolfgang MikulitsLiver sinusoidal endothelial cells (LSECs) represent a highly differentiated cell type that lines hepatic sinusoids. LSECs form a discontinuous endothelium due to fenestrations under physiological conditions, which are reduced upon chronic liver injury. Cultivation of rodent LSECs associates with a rapid onset of stress-induced senescence a few days post isolation, which limits genetic and biochemical studies ex vivo. Here we show the establishment of LSECs isolated from p19ARF-/- mice which undergo more than 50 cell doublings in the absence of senescence. Isolated p19ARF-/- LSECs display a cobblestone-like morphology and show the ability of tube formation. Analysis of DNA content revealed a stable diploid phenotype after long-term passaging without a gain of aneuploidy. Notably, p19ARF-/- LSECs express the endothelial markers CD31, vascular endothelial growth factor receptor (VEGFR)-2, VE-cadherin, von Willebrand factor, stabilin-2 and CD146 suggesting that these cells harbor and maintain an endothelial phenotype. In line, treatment with small molecule inhibitors against VEGFR-2 caused cell death, demonstrating the sustained ability of p19ARF-/- LSECs to respond to anti-angiogenic therapeutics. From these data we conclude that loss of p19ARF overcomes senescence of LSECs, allowing immortalization of cells without losing endothelial characteristics. Thus, p19ARF-/- LSECs provide a novel cellular model to study endothelial cell biology.http://europepmc.org/articles/PMC4631446?pdf=render |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Petra Koudelkova Gerhard Weber Wolfgang Mikulits |
spellingShingle |
Petra Koudelkova Gerhard Weber Wolfgang Mikulits Liver Sinusoidal Endothelial Cells Escape Senescence by Loss of p19ARF. PLoS ONE |
author_facet |
Petra Koudelkova Gerhard Weber Wolfgang Mikulits |
author_sort |
Petra Koudelkova |
title |
Liver Sinusoidal Endothelial Cells Escape Senescence by Loss of p19ARF. |
title_short |
Liver Sinusoidal Endothelial Cells Escape Senescence by Loss of p19ARF. |
title_full |
Liver Sinusoidal Endothelial Cells Escape Senescence by Loss of p19ARF. |
title_fullStr |
Liver Sinusoidal Endothelial Cells Escape Senescence by Loss of p19ARF. |
title_full_unstemmed |
Liver Sinusoidal Endothelial Cells Escape Senescence by Loss of p19ARF. |
title_sort |
liver sinusoidal endothelial cells escape senescence by loss of p19arf. |
publisher |
Public Library of Science (PLoS) |
series |
PLoS ONE |
issn |
1932-6203 |
publishDate |
2015-01-01 |
description |
Liver sinusoidal endothelial cells (LSECs) represent a highly differentiated cell type that lines hepatic sinusoids. LSECs form a discontinuous endothelium due to fenestrations under physiological conditions, which are reduced upon chronic liver injury. Cultivation of rodent LSECs associates with a rapid onset of stress-induced senescence a few days post isolation, which limits genetic and biochemical studies ex vivo. Here we show the establishment of LSECs isolated from p19ARF-/- mice which undergo more than 50 cell doublings in the absence of senescence. Isolated p19ARF-/- LSECs display a cobblestone-like morphology and show the ability of tube formation. Analysis of DNA content revealed a stable diploid phenotype after long-term passaging without a gain of aneuploidy. Notably, p19ARF-/- LSECs express the endothelial markers CD31, vascular endothelial growth factor receptor (VEGFR)-2, VE-cadherin, von Willebrand factor, stabilin-2 and CD146 suggesting that these cells harbor and maintain an endothelial phenotype. In line, treatment with small molecule inhibitors against VEGFR-2 caused cell death, demonstrating the sustained ability of p19ARF-/- LSECs to respond to anti-angiogenic therapeutics. From these data we conclude that loss of p19ARF overcomes senescence of LSECs, allowing immortalization of cells without losing endothelial characteristics. Thus, p19ARF-/- LSECs provide a novel cellular model to study endothelial cell biology. |
url |
http://europepmc.org/articles/PMC4631446?pdf=render |
work_keys_str_mv |
AT petrakoudelkova liversinusoidalendothelialcellsescapesenescencebylossofp19arf AT gerhardweber liversinusoidalendothelialcellsescapesenescencebylossofp19arf AT wolfgangmikulits liversinusoidalendothelialcellsescapesenescencebylossofp19arf |
_version_ |
1725368033623408640 |