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Induction of Human iPSC-Derived Cardiomyocyte Proliferation Revealed by Combinatorial Screening in High Density Microbioreactor Arrays

Inducing cardiomyocyte proliferation in post-mitotic adult heart tissue is attracting significant attention as a therapeutic strategy to regenerate the heart after injury. Model animal screens have identified several candidate signalling pathways, however, it remains unclear as to what extent these...

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Autores principales: Titmarsh, Drew M., Glass, Nick R., Mills, Richard J., Hidalgo, Alejandro, Wolvetang, Ernst J., Porrello, Enzo R., Hudson, James E., Cooper-White, Justin J.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4838928/
https://www.ncbi.nlm.nih.gov/pubmed/27097795
http://dx.doi.org/10.1038/srep24637
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author Titmarsh, Drew M.
Glass, Nick R.
Mills, Richard J.
Hidalgo, Alejandro
Wolvetang, Ernst J.
Porrello, Enzo R.
Hudson, James E.
Cooper-White, Justin J.
author_facet Titmarsh, Drew M.
Glass, Nick R.
Mills, Richard J.
Hidalgo, Alejandro
Wolvetang, Ernst J.
Porrello, Enzo R.
Hudson, James E.
Cooper-White, Justin J.
author_sort Titmarsh, Drew M.
collection PubMed
description Inducing cardiomyocyte proliferation in post-mitotic adult heart tissue is attracting significant attention as a therapeutic strategy to regenerate the heart after injury. Model animal screens have identified several candidate signalling pathways, however, it remains unclear as to what extent these pathways can be exploited, either individually or in combination, in the human system. The advent of human cardiac cells from directed differentiation of human pluripotent stem cells (hPSCs) now provides the ability to interrogate human cardiac biology in vitro, but it remains difficult with existing culture formats to simply and rapidly elucidate signalling pathway penetrance and interplay. To facilitate high-throughput combinatorial screening of candidate biologicals or factors driving relevant molecular pathways, we developed a high-density microbioreactor array (HDMA) – a microfluidic cell culture array containing 8100 culture chambers. We used HDMAs to combinatorially screen Wnt, Hedgehog, IGF and FGF pathway agonists. The Wnt activator CHIR99021 was identified as the most potent molecular inducer of human cardiomyocyte proliferation, inducing cell cycle activity marked by Ki67, and an increase in cardiomyocyte numbers compared to controls. The combination of human cardiomyocytes with the HDMA provides a versatile and rapid tool for stratifying combinations of factors for heart regeneration.
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spelling pubmed-48389282016-04-27 Induction of Human iPSC-Derived Cardiomyocyte Proliferation Revealed by Combinatorial Screening in High Density Microbioreactor Arrays Titmarsh, Drew M. Glass, Nick R. Mills, Richard J. Hidalgo, Alejandro Wolvetang, Ernst J. Porrello, Enzo R. Hudson, James E. Cooper-White, Justin J. Sci Rep Article Inducing cardiomyocyte proliferation in post-mitotic adult heart tissue is attracting significant attention as a therapeutic strategy to regenerate the heart after injury. Model animal screens have identified several candidate signalling pathways, however, it remains unclear as to what extent these pathways can be exploited, either individually or in combination, in the human system. The advent of human cardiac cells from directed differentiation of human pluripotent stem cells (hPSCs) now provides the ability to interrogate human cardiac biology in vitro, but it remains difficult with existing culture formats to simply and rapidly elucidate signalling pathway penetrance and interplay. To facilitate high-throughput combinatorial screening of candidate biologicals or factors driving relevant molecular pathways, we developed a high-density microbioreactor array (HDMA) – a microfluidic cell culture array containing 8100 culture chambers. We used HDMAs to combinatorially screen Wnt, Hedgehog, IGF and FGF pathway agonists. The Wnt activator CHIR99021 was identified as the most potent molecular inducer of human cardiomyocyte proliferation, inducing cell cycle activity marked by Ki67, and an increase in cardiomyocyte numbers compared to controls. The combination of human cardiomyocytes with the HDMA provides a versatile and rapid tool for stratifying combinations of factors for heart regeneration. Nature Publishing Group 2016-04-21 /pmc/articles/PMC4838928/ /pubmed/27097795 http://dx.doi.org/10.1038/srep24637 Text en Copyright © 2016, Macmillan Publishers Limited http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/
spellingShingle Article
Titmarsh, Drew M.
Glass, Nick R.
Mills, Richard J.
Hidalgo, Alejandro
Wolvetang, Ernst J.
Porrello, Enzo R.
Hudson, James E.
Cooper-White, Justin J.
Induction of Human iPSC-Derived Cardiomyocyte Proliferation Revealed by Combinatorial Screening in High Density Microbioreactor Arrays
title Induction of Human iPSC-Derived Cardiomyocyte Proliferation Revealed by Combinatorial Screening in High Density Microbioreactor Arrays
title_full Induction of Human iPSC-Derived Cardiomyocyte Proliferation Revealed by Combinatorial Screening in High Density Microbioreactor Arrays
title_fullStr Induction of Human iPSC-Derived Cardiomyocyte Proliferation Revealed by Combinatorial Screening in High Density Microbioreactor Arrays
title_full_unstemmed Induction of Human iPSC-Derived Cardiomyocyte Proliferation Revealed by Combinatorial Screening in High Density Microbioreactor Arrays
title_short Induction of Human iPSC-Derived Cardiomyocyte Proliferation Revealed by Combinatorial Screening in High Density Microbioreactor Arrays
title_sort induction of human ipsc-derived cardiomyocyte proliferation revealed by combinatorial screening in high density microbioreactor arrays
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4838928/
https://www.ncbi.nlm.nih.gov/pubmed/27097795
http://dx.doi.org/10.1038/srep24637
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