Enhanced proliferation of monolayer cultures of embryonic stem (ES) cell-derived cardiomyocytes following acute loss of retinoblastoma.

<h4>Background</h4>Cardiomyocyte (CM) cell cycle analysis has been impeded because of a reliance on primary neonatal cultures of poorly proliferating cells or chronic transgenic animal models with innate compensatory mechanisms.<h4>Methodology/principal findings</h4>We descri...

Descripción completa

Guardado en:

Detalles Bibliográficos
Autores principales:	Satoshi Yamanaka, Ihor Zahanich, Robert P Wersto, Kenneth R Boheler
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2008
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/fb31032101414b809e089c355c2e9375
Etiquetas:	Agregar Etiqueta Sin Etiquetas, Sea el primero en etiquetar este registro!

id	oai:doaj.org-article:fb31032101414b809e089c355c2e9375
record_format	dspace
spelling	oai:doaj.org-article:fb31032101414b809e089c355c2e93752021-11-25T06:18:14ZEnhanced proliferation of monolayer cultures of embryonic stem (ES) cell-derived cardiomyocytes following acute loss of retinoblastoma.1932-620310.1371/journal.pone.0003896https://doaj.org/article/fb31032101414b809e089c355c2e93752008-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/19066628/pdf/?tool=EBIhttps://doaj.org/toc/1932-6203<h4>Background</h4>Cardiomyocyte (CM) cell cycle analysis has been impeded because of a reliance on primary neonatal cultures of poorly proliferating cells or chronic transgenic animal models with innate compensatory mechanisms.<h4>Methodology/principal findings</h4>We describe an in vitro model consisting of monolayer cultures of highly proliferative embryonic stem (ES) cell-derived CM. Following induction with ascorbate and selection with puromycin, early CM cultures are >98% pure, and at least 85% of the cells actively proliferate. During the proliferative stage, cells express high levels of E2F3a, B-Myb and phosphorylated forms of retinoblastoma (Rb), but with continued cultivation, cells stop dividing and mature functionally. This developmental transition is characterized by a switch from slow skeletal to cardiac TnI, an increase in binucleation, cardiac calsequestrin and hypophosphorylated Rb, a decrease in E2F3, B-Myb and atrial natriuretic factor, and the establishment of a more negative resting membrane potential. Although previous publications suggested that Rb was not necessary for cell cycle control in heart, we find following acute knockdown of Rb that this factor actively regulates progression through the G1 checkpoint and that its loss promotes proliferation at the expense of CM maturation.<h4>Conclusions/significance</h4>We have established a unique model system for studying cardiac cell cycle progression, and show in contrast to previous reports that Rb actively regulates both cell cycle progression through the G1 checkpoint and maturation of heart cells. We conclude that this in vitro model will facilitate the analysis of cell cycle control mechanisms of CMs.Satoshi YamanakaIhor ZahanichRobert P WerstoKenneth R BohelerPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 3, Iss 12, p e3896 (2008)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Satoshi Yamanaka Ihor Zahanich Robert P Wersto Kenneth R Boheler Enhanced proliferation of monolayer cultures of embryonic stem (ES) cell-derived cardiomyocytes following acute loss of retinoblastoma.
description	<h4>Background</h4>Cardiomyocyte (CM) cell cycle analysis has been impeded because of a reliance on primary neonatal cultures of poorly proliferating cells or chronic transgenic animal models with innate compensatory mechanisms.<h4>Methodology/principal findings</h4>We describe an in vitro model consisting of monolayer cultures of highly proliferative embryonic stem (ES) cell-derived CM. Following induction with ascorbate and selection with puromycin, early CM cultures are >98% pure, and at least 85% of the cells actively proliferate. During the proliferative stage, cells express high levels of E2F3a, B-Myb and phosphorylated forms of retinoblastoma (Rb), but with continued cultivation, cells stop dividing and mature functionally. This developmental transition is characterized by a switch from slow skeletal to cardiac TnI, an increase in binucleation, cardiac calsequestrin and hypophosphorylated Rb, a decrease in E2F3, B-Myb and atrial natriuretic factor, and the establishment of a more negative resting membrane potential. Although previous publications suggested that Rb was not necessary for cell cycle control in heart, we find following acute knockdown of Rb that this factor actively regulates progression through the G1 checkpoint and that its loss promotes proliferation at the expense of CM maturation.<h4>Conclusions/significance</h4>We have established a unique model system for studying cardiac cell cycle progression, and show in contrast to previous reports that Rb actively regulates both cell cycle progression through the G1 checkpoint and maturation of heart cells. We conclude that this in vitro model will facilitate the analysis of cell cycle control mechanisms of CMs.
format	article
author	Satoshi Yamanaka Ihor Zahanich Robert P Wersto Kenneth R Boheler
author_facet	Satoshi Yamanaka Ihor Zahanich Robert P Wersto Kenneth R Boheler
author_sort	Satoshi Yamanaka
title	Enhanced proliferation of monolayer cultures of embryonic stem (ES) cell-derived cardiomyocytes following acute loss of retinoblastoma.
title_short	Enhanced proliferation of monolayer cultures of embryonic stem (ES) cell-derived cardiomyocytes following acute loss of retinoblastoma.
title_full	Enhanced proliferation of monolayer cultures of embryonic stem (ES) cell-derived cardiomyocytes following acute loss of retinoblastoma.
title_fullStr	Enhanced proliferation of monolayer cultures of embryonic stem (ES) cell-derived cardiomyocytes following acute loss of retinoblastoma.
title_full_unstemmed	Enhanced proliferation of monolayer cultures of embryonic stem (ES) cell-derived cardiomyocytes following acute loss of retinoblastoma.
title_sort	enhanced proliferation of monolayer cultures of embryonic stem (es) cell-derived cardiomyocytes following acute loss of retinoblastoma.
publisher	Public Library of Science (PLoS)
publishDate	2008
url	https://doaj.org/article/fb31032101414b809e089c355c2e9375
work_keys_str_mv	AT satoshiyamanaka enhancedproliferationofmonolayerculturesofembryonicstemescellderivedcardiomyocytesfollowingacutelossofretinoblastoma AT ihorzahanich enhancedproliferationofmonolayerculturesofembryonicstemescellderivedcardiomyocytesfollowingacutelossofretinoblastoma AT robertpwersto enhancedproliferationofmonolayerculturesofembryonicstemescellderivedcardiomyocytesfollowingacutelossofretinoblastoma AT kennethrboheler enhancedproliferationofmonolayerculturesofembryonicstemescellderivedcardiomyocytesfollowingacutelossofretinoblastoma
_version_	1718413961808838656

Enhanced proliferation of monolayer cultures of embryonic stem (ES) cell-derived cardiomyocytes following acute loss of retinoblastoma.

Ejemplares similares