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Isolation and Characterization of Primary Rat Valve Interstitial Cells: A New Model to Study Aortic Valve Calcification

Calcific aortic valve disease (CAVD) is characterized by the progressive thickening of the aortic valve leaflets. It is a condition frequently found in the elderly and end-stage renal disease (ESRD) patients, who commonly suffer from hyperphosphatemia and hypercalcemia. At present, there are no medi...

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Autores principales: Lin, Cui, Zhu, Dongxing, Markby, Greg, Corcoran, Brendan M., Farquharson, Colin, Macrae, Vicky E.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MyJove Corporation 2017
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5755457/
https://www.ncbi.nlm.nih.gov/pubmed/29286439
http://dx.doi.org/10.3791/56126
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author Lin, Cui
Zhu, Dongxing
Markby, Greg
Corcoran, Brendan M.
Farquharson, Colin
Macrae, Vicky E.
author_facet Lin, Cui
Zhu, Dongxing
Markby, Greg
Corcoran, Brendan M.
Farquharson, Colin
Macrae, Vicky E.
author_sort Lin, Cui
collection PubMed
description Calcific aortic valve disease (CAVD) is characterized by the progressive thickening of the aortic valve leaflets. It is a condition frequently found in the elderly and end-stage renal disease (ESRD) patients, who commonly suffer from hyperphosphatemia and hypercalcemia. At present, there are no medication therapies that can stop its progression. The mechanisms that underlie this pathological process remain unclear. The aortic valve leaflet is composed of a thin layer of valve endothelial cells (VECs) on the outer surfaces of the aortic cusps, with valve interstitial cells (VICs) sandwiched between the VECs. The use of a rat model enables the in vitro study of ectopic calcification based on the in vivo physiopathological serum phosphate (Pi) and calcium (Ca) levels of patients who suffer from hyperphosphatemia and hypercalcemia. The described protocol details the isolation of a pure rat VIC population as shown by the expression of VIC markers: alpha-smooth muscle actin (α-SMA) vimentin and tissue growth factor beta (TGFβ) 1 and 2, and the absence of cluster of differentiation (CD) 31, a VEC marker. By expanding these VICs, biochemical, genetic, and imaging studies can be performed to study and unravel the key mediators underpinning CAVD.
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spelling pubmed-57554572018-01-19 Isolation and Characterization of Primary Rat Valve Interstitial Cells: A New Model to Study Aortic Valve Calcification Lin, Cui Zhu, Dongxing Markby, Greg Corcoran, Brendan M. Farquharson, Colin Macrae, Vicky E. J Vis Exp Developmental Biology Calcific aortic valve disease (CAVD) is characterized by the progressive thickening of the aortic valve leaflets. It is a condition frequently found in the elderly and end-stage renal disease (ESRD) patients, who commonly suffer from hyperphosphatemia and hypercalcemia. At present, there are no medication therapies that can stop its progression. The mechanisms that underlie this pathological process remain unclear. The aortic valve leaflet is composed of a thin layer of valve endothelial cells (VECs) on the outer surfaces of the aortic cusps, with valve interstitial cells (VICs) sandwiched between the VECs. The use of a rat model enables the in vitro study of ectopic calcification based on the in vivo physiopathological serum phosphate (Pi) and calcium (Ca) levels of patients who suffer from hyperphosphatemia and hypercalcemia. The described protocol details the isolation of a pure rat VIC population as shown by the expression of VIC markers: alpha-smooth muscle actin (α-SMA) vimentin and tissue growth factor beta (TGFβ) 1 and 2, and the absence of cluster of differentiation (CD) 31, a VEC marker. By expanding these VICs, biochemical, genetic, and imaging studies can be performed to study and unravel the key mediators underpinning CAVD. MyJove Corporation 2017-11-20 /pmc/articles/PMC5755457/ /pubmed/29286439 http://dx.doi.org/10.3791/56126 Text en Copyright © 2017, Journal of Visualized Experiments http://creativecommons.org/licenses/by/3.0/us/ This is an open-access article distributed under the terms of the Creative Commons Attribution 3.0 License. To view a copy of this license, visithttp://creativecommons.org/licenses/by/3.0/us/
spellingShingle Developmental Biology
Lin, Cui
Zhu, Dongxing
Markby, Greg
Corcoran, Brendan M.
Farquharson, Colin
Macrae, Vicky E.
Isolation and Characterization of Primary Rat Valve Interstitial Cells: A New Model to Study Aortic Valve Calcification
title Isolation and Characterization of Primary Rat Valve Interstitial Cells: A New Model to Study Aortic Valve Calcification
title_full Isolation and Characterization of Primary Rat Valve Interstitial Cells: A New Model to Study Aortic Valve Calcification
title_fullStr Isolation and Characterization of Primary Rat Valve Interstitial Cells: A New Model to Study Aortic Valve Calcification
title_full_unstemmed Isolation and Characterization of Primary Rat Valve Interstitial Cells: A New Model to Study Aortic Valve Calcification
title_short Isolation and Characterization of Primary Rat Valve Interstitial Cells: A New Model to Study Aortic Valve Calcification
title_sort isolation and characterization of primary rat valve interstitial cells: a new model to study aortic valve calcification
topic Developmental Biology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5755457/
https://www.ncbi.nlm.nih.gov/pubmed/29286439
http://dx.doi.org/10.3791/56126
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