Cargando…

Co-culture with neonatal cardiomyocytes enhances the proliferation of iPSC-derived cardiomyocytes via FAK/JNK signaling

BACKGROUND: We previously reported that the pluripotent stem cells can differentiate into cardiomyocytes (CMs) by co-culture with neonatal CMs (NCMs) in vitro. However, the involving mechanism is not clear. METHODS: Mouse induced pluripotent stem cells (iPSCs) were cultured in hanging drops to form...

Descripción completa

Detalles Bibliográficos
Autores principales: Ou, Dongbo, Wang, Qi, Huang, Yanjin, Zeng, Di, Wei, Ting, Ding, Lu, Li, Xiaoli, Zheng, Qiangsun, Jin, Yan
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2016
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4855360/
https://www.ncbi.nlm.nih.gov/pubmed/27141946
http://dx.doi.org/10.1186/s12861-016-0112-2
_version_ 1782430354730123264
author Ou, Dongbo
Wang, Qi
Huang, Yanjin
Zeng, Di
Wei, Ting
Ding, Lu
Li, Xiaoli
Zheng, Qiangsun
Jin, Yan
author_facet Ou, Dongbo
Wang, Qi
Huang, Yanjin
Zeng, Di
Wei, Ting
Ding, Lu
Li, Xiaoli
Zheng, Qiangsun
Jin, Yan
author_sort Ou, Dongbo
collection PubMed
description BACKGROUND: We previously reported that the pluripotent stem cells can differentiate into cardiomyocytes (CMs) by co-culture with neonatal CMs (NCMs) in vitro. However, the involving mechanism is not clear. METHODS: Mouse induced pluripotent stem cells (iPSCs) were cultured in hanging drops to form embryoid bodies (EBs) and to induce myocardial differentiation. Co-culture of EBs and NCMs was established in a transwell insert system, while EBs grown alone in the wells were used as controls. RESULTS: Co-culture with NCMs markedly increased the generation of functional CMs from iPSCs. The focal adhesion kinase (FAK) phosphorylation, and c-Jun N-terminal kinase (JNK) phosphorylation in co-culture were higher than that in EBs grown alone. Treating FAK small interfering RNA (FAK siRNA) or specific inhibitor for JNK (SP600125) to iPSCs significantly reduced the phosphorylation of JNK and the expressions of Mef2c and Bcl-2. The expressions of cTnT and MLC-2V were also decreased. Our results revealed that co-culture with NCMs significantly enhance the differentiation ability of iPSCs by increasing Mef2c and Bcl-2 expressions concomitantly with a marked augment on cell proliferation through JNK signaling pathways. CONCLUSIONS: These findings indicated that co-culture of EBs with NCMs induces genes expressed in a mature pattern and stimulates the proliferation of iPSC-derived CMs (iPS-CMs) by activating FAK/JNK signaling. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12861-016-0112-2) contains supplementary material, which is available to authorized users.
format Online
Article
Text
id pubmed-4855360
institution National Center for Biotechnology Information
language English
publishDate 2016
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-48553602016-05-05 Co-culture with neonatal cardiomyocytes enhances the proliferation of iPSC-derived cardiomyocytes via FAK/JNK signaling Ou, Dongbo Wang, Qi Huang, Yanjin Zeng, Di Wei, Ting Ding, Lu Li, Xiaoli Zheng, Qiangsun Jin, Yan BMC Dev Biol Research Article BACKGROUND: We previously reported that the pluripotent stem cells can differentiate into cardiomyocytes (CMs) by co-culture with neonatal CMs (NCMs) in vitro. However, the involving mechanism is not clear. METHODS: Mouse induced pluripotent stem cells (iPSCs) were cultured in hanging drops to form embryoid bodies (EBs) and to induce myocardial differentiation. Co-culture of EBs and NCMs was established in a transwell insert system, while EBs grown alone in the wells were used as controls. RESULTS: Co-culture with NCMs markedly increased the generation of functional CMs from iPSCs. The focal adhesion kinase (FAK) phosphorylation, and c-Jun N-terminal kinase (JNK) phosphorylation in co-culture were higher than that in EBs grown alone. Treating FAK small interfering RNA (FAK siRNA) or specific inhibitor for JNK (SP600125) to iPSCs significantly reduced the phosphorylation of JNK and the expressions of Mef2c and Bcl-2. The expressions of cTnT and MLC-2V were also decreased. Our results revealed that co-culture with NCMs significantly enhance the differentiation ability of iPSCs by increasing Mef2c and Bcl-2 expressions concomitantly with a marked augment on cell proliferation through JNK signaling pathways. CONCLUSIONS: These findings indicated that co-culture of EBs with NCMs induces genes expressed in a mature pattern and stimulates the proliferation of iPSC-derived CMs (iPS-CMs) by activating FAK/JNK signaling. ELECTRONIC SUPPLEMENTARY MATERIAL: The online version of this article (doi:10.1186/s12861-016-0112-2) contains supplementary material, which is available to authorized users. BioMed Central 2016-05-04 /pmc/articles/PMC4855360/ /pubmed/27141946 http://dx.doi.org/10.1186/s12861-016-0112-2 Text en © Ou et al. 2016 Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver (http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.
spellingShingle Research Article
Ou, Dongbo
Wang, Qi
Huang, Yanjin
Zeng, Di
Wei, Ting
Ding, Lu
Li, Xiaoli
Zheng, Qiangsun
Jin, Yan
Co-culture with neonatal cardiomyocytes enhances the proliferation of iPSC-derived cardiomyocytes via FAK/JNK signaling
title Co-culture with neonatal cardiomyocytes enhances the proliferation of iPSC-derived cardiomyocytes via FAK/JNK signaling
title_full Co-culture with neonatal cardiomyocytes enhances the proliferation of iPSC-derived cardiomyocytes via FAK/JNK signaling
title_fullStr Co-culture with neonatal cardiomyocytes enhances the proliferation of iPSC-derived cardiomyocytes via FAK/JNK signaling
title_full_unstemmed Co-culture with neonatal cardiomyocytes enhances the proliferation of iPSC-derived cardiomyocytes via FAK/JNK signaling
title_short Co-culture with neonatal cardiomyocytes enhances the proliferation of iPSC-derived cardiomyocytes via FAK/JNK signaling
title_sort co-culture with neonatal cardiomyocytes enhances the proliferation of ipsc-derived cardiomyocytes via fak/jnk signaling
topic Research Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4855360/
https://www.ncbi.nlm.nih.gov/pubmed/27141946
http://dx.doi.org/10.1186/s12861-016-0112-2
work_keys_str_mv AT oudongbo coculturewithneonatalcardiomyocytesenhancestheproliferationofipscderivedcardiomyocytesviafakjnksignaling
AT wangqi coculturewithneonatalcardiomyocytesenhancestheproliferationofipscderivedcardiomyocytesviafakjnksignaling
AT huangyanjin coculturewithneonatalcardiomyocytesenhancestheproliferationofipscderivedcardiomyocytesviafakjnksignaling
AT zengdi coculturewithneonatalcardiomyocytesenhancestheproliferationofipscderivedcardiomyocytesviafakjnksignaling
AT weiting coculturewithneonatalcardiomyocytesenhancestheproliferationofipscderivedcardiomyocytesviafakjnksignaling
AT dinglu coculturewithneonatalcardiomyocytesenhancestheproliferationofipscderivedcardiomyocytesviafakjnksignaling
AT lixiaoli coculturewithneonatalcardiomyocytesenhancestheproliferationofipscderivedcardiomyocytesviafakjnksignaling
AT zhengqiangsun coculturewithneonatalcardiomyocytesenhancestheproliferationofipscderivedcardiomyocytesviafakjnksignaling
AT jinyan coculturewithneonatalcardiomyocytesenhancestheproliferationofipscderivedcardiomyocytesviafakjnksignaling