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Unraveling the Inconsistencies of Cardiac Differentiation Efficiency Induced by the GSK3β Inhibitor CHIR99021 in Human Pluripotent Stem Cells
Cardiac differentiation efficiency is hampered by inconsistencies and low reproducibility. We analyzed the differentiation process of multiple human pluripotent stem cell (hPSC) lines in response to dynamic GSK3β inhibition under varying cell culture conditions. hPSCs showed strong differences in ce...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Elsevier
2018
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5989659/ https://www.ncbi.nlm.nih.gov/pubmed/29706502 http://dx.doi.org/10.1016/j.stemcr.2018.03.023 |
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author | Laco, Filip Woo, Tsung Liang Zhong, Qixing Szmyd, Radoslaw Ting, Sherwin Khan, Fahima Jaleel Chai, Christina L.L. Reuveny, Shaul Chen, Allen Oh, Steve |
author_facet | Laco, Filip Woo, Tsung Liang Zhong, Qixing Szmyd, Radoslaw Ting, Sherwin Khan, Fahima Jaleel Chai, Christina L.L. Reuveny, Shaul Chen, Allen Oh, Steve |
author_sort | Laco, Filip |
collection | PubMed |
description | Cardiac differentiation efficiency is hampered by inconsistencies and low reproducibility. We analyzed the differentiation process of multiple human pluripotent stem cell (hPSC) lines in response to dynamic GSK3β inhibition under varying cell culture conditions. hPSCs showed strong differences in cell-cycle profiles with varying culture confluency. hPSCs with a higher percentage of cells in the G1 phase of the cell cycle exhibited cell death and required lower doses of GSK3β inhibitors to induce cardiac differentiation. GSK3β inhibition initiated cell-cycle progression via cyclin D1 and modulated both Wnt signaling and the transcription factor (TCF) levels, resulting in accelerated or delayed mesoderm differentiation. The TCF levels were key regulators during hPSC differentiation with CHIR99021. Our results explain how differences in hPSC lines and culture conditions impact cell death and cardiac differentiation. By analyzing the cell cycle, we were able to select for highly cardiogenic hPSC lines and increase the experimental reproducibility by predicting differentiation outcomes. |
format | Online Article Text |
id | pubmed-5989659 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2018 |
publisher | Elsevier |
record_format | MEDLINE/PubMed |
spelling | pubmed-59896592018-06-07 Unraveling the Inconsistencies of Cardiac Differentiation Efficiency Induced by the GSK3β Inhibitor CHIR99021 in Human Pluripotent Stem Cells Laco, Filip Woo, Tsung Liang Zhong, Qixing Szmyd, Radoslaw Ting, Sherwin Khan, Fahima Jaleel Chai, Christina L.L. Reuveny, Shaul Chen, Allen Oh, Steve Stem Cell Reports Article Cardiac differentiation efficiency is hampered by inconsistencies and low reproducibility. We analyzed the differentiation process of multiple human pluripotent stem cell (hPSC) lines in response to dynamic GSK3β inhibition under varying cell culture conditions. hPSCs showed strong differences in cell-cycle profiles with varying culture confluency. hPSCs with a higher percentage of cells in the G1 phase of the cell cycle exhibited cell death and required lower doses of GSK3β inhibitors to induce cardiac differentiation. GSK3β inhibition initiated cell-cycle progression via cyclin D1 and modulated both Wnt signaling and the transcription factor (TCF) levels, resulting in accelerated or delayed mesoderm differentiation. The TCF levels were key regulators during hPSC differentiation with CHIR99021. Our results explain how differences in hPSC lines and culture conditions impact cell death and cardiac differentiation. By analyzing the cell cycle, we were able to select for highly cardiogenic hPSC lines and increase the experimental reproducibility by predicting differentiation outcomes. Elsevier 2018-04-26 /pmc/articles/PMC5989659/ /pubmed/29706502 http://dx.doi.org/10.1016/j.stemcr.2018.03.023 Text en © 2018 The Authors http://creativecommons.org/licenses/by/4.0/ This is an open access article under the CC BY license (http://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Laco, Filip Woo, Tsung Liang Zhong, Qixing Szmyd, Radoslaw Ting, Sherwin Khan, Fahima Jaleel Chai, Christina L.L. Reuveny, Shaul Chen, Allen Oh, Steve Unraveling the Inconsistencies of Cardiac Differentiation Efficiency Induced by the GSK3β Inhibitor CHIR99021 in Human Pluripotent Stem Cells |
title | Unraveling the Inconsistencies of Cardiac Differentiation Efficiency Induced by the GSK3β Inhibitor CHIR99021 in Human Pluripotent Stem Cells |
title_full | Unraveling the Inconsistencies of Cardiac Differentiation Efficiency Induced by the GSK3β Inhibitor CHIR99021 in Human Pluripotent Stem Cells |
title_fullStr | Unraveling the Inconsistencies of Cardiac Differentiation Efficiency Induced by the GSK3β Inhibitor CHIR99021 in Human Pluripotent Stem Cells |
title_full_unstemmed | Unraveling the Inconsistencies of Cardiac Differentiation Efficiency Induced by the GSK3β Inhibitor CHIR99021 in Human Pluripotent Stem Cells |
title_short | Unraveling the Inconsistencies of Cardiac Differentiation Efficiency Induced by the GSK3β Inhibitor CHIR99021 in Human Pluripotent Stem Cells |
title_sort | unraveling the inconsistencies of cardiac differentiation efficiency induced by the gsk3β inhibitor chir99021 in human pluripotent stem cells |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5989659/ https://www.ncbi.nlm.nih.gov/pubmed/29706502 http://dx.doi.org/10.1016/j.stemcr.2018.03.023 |
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