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Cardiac differentiation of human pluripotent stem cells using defined extracellular matrix proteins reveals essential role of fibronectin
Research and therapeutic applications using human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) require robust differentiation strategies. Efforts to improve hPSC-CM differentiation have largely overlooked the role of extracellular matrix (ECM). The present study investigates the ability o...
Autores principales: | , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
eLife Sciences Publications, Ltd
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9236614/ https://www.ncbi.nlm.nih.gov/pubmed/35758861 http://dx.doi.org/10.7554/eLife.69028 |
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author | Zhang, Jianhua Gregorich, Zachery R Tao, Ran Kim, Gina C Lalit, Pratik A Carvalho, Juliana L Markandeya, Yogananda Mosher, Deane F Palecek, Sean P Kamp, Timothy J |
author_facet | Zhang, Jianhua Gregorich, Zachery R Tao, Ran Kim, Gina C Lalit, Pratik A Carvalho, Juliana L Markandeya, Yogananda Mosher, Deane F Palecek, Sean P Kamp, Timothy J |
author_sort | Zhang, Jianhua |
collection | PubMed |
description | Research and therapeutic applications using human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) require robust differentiation strategies. Efforts to improve hPSC-CM differentiation have largely overlooked the role of extracellular matrix (ECM). The present study investigates the ability of defined ECM proteins to promote hPSC cardiac differentiation. Fibronectin (FN), laminin-111, and laminin-521 enabled hPSCs to attach and expand. However, only addition of FN promoted cardiac differentiation in response to growth factors Activin A, BMP4, and bFGF in contrast to the inhibition produced by laminin-111 or laminin-521. hPSCs in culture produced endogenous FN which accumulated in the ECM to a critical level necessary for effective cardiac differentiation. Inducible shRNA knockdown of FN prevented Brachyury(+) mesoderm formation and subsequent hPSC-CM generation. Antibodies blocking FN binding integrins α4β1 or αVβ1, but not α5β1, inhibited cardiac differentiation. Furthermore, inhibition of integrin-linked kinase led to a decrease in phosphorylated AKT, which was associated with increased apoptosis and inhibition of cardiac differentiation. These results provide new insights into defined matrices for culture of hPSCs that enable production of FN-enriched ECM which is essential for mesoderm formation and efficient cardiac differentiation. |
format | Online Article Text |
id | pubmed-9236614 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2022 |
publisher | eLife Sciences Publications, Ltd |
record_format | MEDLINE/PubMed |
spelling | pubmed-92366142022-06-28 Cardiac differentiation of human pluripotent stem cells using defined extracellular matrix proteins reveals essential role of fibronectin Zhang, Jianhua Gregorich, Zachery R Tao, Ran Kim, Gina C Lalit, Pratik A Carvalho, Juliana L Markandeya, Yogananda Mosher, Deane F Palecek, Sean P Kamp, Timothy J eLife Stem Cells and Regenerative Medicine Research and therapeutic applications using human pluripotent stem cell-derived cardiomyocytes (hPSC-CMs) require robust differentiation strategies. Efforts to improve hPSC-CM differentiation have largely overlooked the role of extracellular matrix (ECM). The present study investigates the ability of defined ECM proteins to promote hPSC cardiac differentiation. Fibronectin (FN), laminin-111, and laminin-521 enabled hPSCs to attach and expand. However, only addition of FN promoted cardiac differentiation in response to growth factors Activin A, BMP4, and bFGF in contrast to the inhibition produced by laminin-111 or laminin-521. hPSCs in culture produced endogenous FN which accumulated in the ECM to a critical level necessary for effective cardiac differentiation. Inducible shRNA knockdown of FN prevented Brachyury(+) mesoderm formation and subsequent hPSC-CM generation. Antibodies blocking FN binding integrins α4β1 or αVβ1, but not α5β1, inhibited cardiac differentiation. Furthermore, inhibition of integrin-linked kinase led to a decrease in phosphorylated AKT, which was associated with increased apoptosis and inhibition of cardiac differentiation. These results provide new insights into defined matrices for culture of hPSCs that enable production of FN-enriched ECM which is essential for mesoderm formation and efficient cardiac differentiation. eLife Sciences Publications, Ltd 2022-06-27 /pmc/articles/PMC9236614/ /pubmed/35758861 http://dx.doi.org/10.7554/eLife.69028 Text en © 2022, Zhang et al https://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (https://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited. |
spellingShingle | Stem Cells and Regenerative Medicine Zhang, Jianhua Gregorich, Zachery R Tao, Ran Kim, Gina C Lalit, Pratik A Carvalho, Juliana L Markandeya, Yogananda Mosher, Deane F Palecek, Sean P Kamp, Timothy J Cardiac differentiation of human pluripotent stem cells using defined extracellular matrix proteins reveals essential role of fibronectin |
title | Cardiac differentiation of human pluripotent stem cells using defined extracellular matrix proteins reveals essential role of fibronectin |
title_full | Cardiac differentiation of human pluripotent stem cells using defined extracellular matrix proteins reveals essential role of fibronectin |
title_fullStr | Cardiac differentiation of human pluripotent stem cells using defined extracellular matrix proteins reveals essential role of fibronectin |
title_full_unstemmed | Cardiac differentiation of human pluripotent stem cells using defined extracellular matrix proteins reveals essential role of fibronectin |
title_short | Cardiac differentiation of human pluripotent stem cells using defined extracellular matrix proteins reveals essential role of fibronectin |
title_sort | cardiac differentiation of human pluripotent stem cells using defined extracellular matrix proteins reveals essential role of fibronectin |
topic | Stem Cells and Regenerative Medicine |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9236614/ https://www.ncbi.nlm.nih.gov/pubmed/35758861 http://dx.doi.org/10.7554/eLife.69028 |
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