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Generation of functional cardiomyocytes from rat embryonic and induced pluripotent stem cells using feeder-free expansion and differentiation in suspension culture

The possibility to generate cardiomyocytes from pluripotent stem cells in vitro has enormous significance for basic research, disease modeling, drug development and heart repair. The concept of heart muscle reconstruction has been studied and optimized in the rat model using rat primary cardiovascul...

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Autores principales: Dahlmann, Julia, Awad, George, Dolny, Carsten, Weinert, Sönke, Richter, Karin, Fischer, Klaus-Dieter, Munsch, Thomas, Leßmann, Volkmar, Volleth, Marianne, Zenker, Martin, Chen, Yaoyao, Merkl, Claudia, Schnieke, Angelika, Baraki, Hassina, Kutschka, Ingo, Kensah, George
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Public Library of Science 2018
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5841662/
https://www.ncbi.nlm.nih.gov/pubmed/29513687
http://dx.doi.org/10.1371/journal.pone.0192652
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author Dahlmann, Julia
Awad, George
Dolny, Carsten
Weinert, Sönke
Richter, Karin
Fischer, Klaus-Dieter
Munsch, Thomas
Leßmann, Volkmar
Volleth, Marianne
Zenker, Martin
Chen, Yaoyao
Merkl, Claudia
Schnieke, Angelika
Baraki, Hassina
Kutschka, Ingo
Kensah, George
author_facet Dahlmann, Julia
Awad, George
Dolny, Carsten
Weinert, Sönke
Richter, Karin
Fischer, Klaus-Dieter
Munsch, Thomas
Leßmann, Volkmar
Volleth, Marianne
Zenker, Martin
Chen, Yaoyao
Merkl, Claudia
Schnieke, Angelika
Baraki, Hassina
Kutschka, Ingo
Kensah, George
author_sort Dahlmann, Julia
collection PubMed
description The possibility to generate cardiomyocytes from pluripotent stem cells in vitro has enormous significance for basic research, disease modeling, drug development and heart repair. The concept of heart muscle reconstruction has been studied and optimized in the rat model using rat primary cardiovascular cells or xenogeneic pluripotent stem cell derived-cardiomyocytes for years. However, the lack of rat pluripotent stem cells (rPSCs) and their cardiovascular derivatives prevented the establishment of an authentic clinically relevant syngeneic or allogeneic rat heart regeneration model. In this study, we comparatively explored the potential of recently available rat embryonic stem cells (rESCs) and induced pluripotent stem cells (riPSCs) as a source for cardiomyocytes (CMs). We developed feeder cell-free culture conditions facilitating the expansion of undifferentiated rPSCs and initiated cardiac differentiation by embryoid body (EB)-formation in agarose microwell arrays, which substituted the robust but labor-intensive hanging drop (HD) method. Ascorbic acid was identified as an efficient enhancer of cardiac differentiation in both rPSC types by significantly increasing the number of beating EBs (3.6 ± 1.6-fold for rESCs and 17.6 ± 3.2-fold for riPSCs). These optimizations resulted in a differentiation efficiency of up to 20% cTnT(pos) rPSC-derived CMs. CMs showed spontaneous contractions, expressed cardiac markers and had typical morphological features. Electrophysiology of riPSC-CMs revealed different cardiac subtypes and physiological responses to cardio-active drugs. In conclusion, we describe rPSCs as a robust source of CMs, which is a prerequisite for detailed preclinical studies of myocardial reconstruction in a physiologically and immunologically relevant small animal model.
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spelling pubmed-58416622018-03-23 Generation of functional cardiomyocytes from rat embryonic and induced pluripotent stem cells using feeder-free expansion and differentiation in suspension culture Dahlmann, Julia Awad, George Dolny, Carsten Weinert, Sönke Richter, Karin Fischer, Klaus-Dieter Munsch, Thomas Leßmann, Volkmar Volleth, Marianne Zenker, Martin Chen, Yaoyao Merkl, Claudia Schnieke, Angelika Baraki, Hassina Kutschka, Ingo Kensah, George PLoS One Research Article The possibility to generate cardiomyocytes from pluripotent stem cells in vitro has enormous significance for basic research, disease modeling, drug development and heart repair. The concept of heart muscle reconstruction has been studied and optimized in the rat model using rat primary cardiovascular cells or xenogeneic pluripotent stem cell derived-cardiomyocytes for years. However, the lack of rat pluripotent stem cells (rPSCs) and their cardiovascular derivatives prevented the establishment of an authentic clinically relevant syngeneic or allogeneic rat heart regeneration model. In this study, we comparatively explored the potential of recently available rat embryonic stem cells (rESCs) and induced pluripotent stem cells (riPSCs) as a source for cardiomyocytes (CMs). We developed feeder cell-free culture conditions facilitating the expansion of undifferentiated rPSCs and initiated cardiac differentiation by embryoid body (EB)-formation in agarose microwell arrays, which substituted the robust but labor-intensive hanging drop (HD) method. Ascorbic acid was identified as an efficient enhancer of cardiac differentiation in both rPSC types by significantly increasing the number of beating EBs (3.6 ± 1.6-fold for rESCs and 17.6 ± 3.2-fold for riPSCs). These optimizations resulted in a differentiation efficiency of up to 20% cTnT(pos) rPSC-derived CMs. CMs showed spontaneous contractions, expressed cardiac markers and had typical morphological features. Electrophysiology of riPSC-CMs revealed different cardiac subtypes and physiological responses to cardio-active drugs. In conclusion, we describe rPSCs as a robust source of CMs, which is a prerequisite for detailed preclinical studies of myocardial reconstruction in a physiologically and immunologically relevant small animal model. Public Library of Science 2018-03-07 /pmc/articles/PMC5841662/ /pubmed/29513687 http://dx.doi.org/10.1371/journal.pone.0192652 Text en © 2018 Dahlmann et al http://creativecommons.org/licenses/by/4.0/ This is an open access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited.
spellingShingle Research Article
Dahlmann, Julia
Awad, George
Dolny, Carsten
Weinert, Sönke
Richter, Karin
Fischer, Klaus-Dieter
Munsch, Thomas
Leßmann, Volkmar
Volleth, Marianne
Zenker, Martin
Chen, Yaoyao
Merkl, Claudia
Schnieke, Angelika
Baraki, Hassina
Kutschka, Ingo
Kensah, George
Generation of functional cardiomyocytes from rat embryonic and induced pluripotent stem cells using feeder-free expansion and differentiation in suspension culture
title Generation of functional cardiomyocytes from rat embryonic and induced pluripotent stem cells using feeder-free expansion and differentiation in suspension culture
title_full Generation of functional cardiomyocytes from rat embryonic and induced pluripotent stem cells using feeder-free expansion and differentiation in suspension culture
title_fullStr Generation of functional cardiomyocytes from rat embryonic and induced pluripotent stem cells using feeder-free expansion and differentiation in suspension culture
title_full_unstemmed Generation of functional cardiomyocytes from rat embryonic and induced pluripotent stem cells using feeder-free expansion and differentiation in suspension culture
title_short Generation of functional cardiomyocytes from rat embryonic and induced pluripotent stem cells using feeder-free expansion and differentiation in suspension culture
title_sort generation of functional cardiomyocytes from rat embryonic and induced pluripotent stem cells using feeder-free expansion and differentiation in suspension culture
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5841662/
https://www.ncbi.nlm.nih.gov/pubmed/29513687
http://dx.doi.org/10.1371/journal.pone.0192652
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