Culture conditions affect cardiac differentiation potential of human pluripotent stem cells.

Human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), are capable of differentiating into any cell type in the human body and thus can be used in studies of early human development, as cell models for different diseases...

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Autores principales:	Marisa Ojala, Kristiina Rajala, Mari Pekkanen-Mattila, Marinka Miettinen, Heini Huhtala, Katriina Aalto-Setälä
Formato:	article
Lenguaje:	EN
Publicado:	Public Library of Science (PLoS) 2012
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/6b05ac8c438c4d21b5ac8017de523eb0
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spelling	oai:doaj.org-article:6b05ac8c438c4d21b5ac8017de523eb02021-11-18T08:10:17ZCulture conditions affect cardiac differentiation potential of human pluripotent stem cells.1932-620310.1371/journal.pone.0048659https://doaj.org/article/6b05ac8c438c4d21b5ac8017de523eb02012-01-01T00:00:00Zhttps://www.ncbi.nlm.nih.gov/pmc/articles/pmid/23119085/?tool=EBIhttps://doaj.org/toc/1932-6203Human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), are capable of differentiating into any cell type in the human body and thus can be used in studies of early human development, as cell models for different diseases and eventually also in regenerative medicine applications. Since the first derivation of hESCs in 1998, a variety of culture conditions have been described for the undifferentiated growth of hPSCs. In this study, we cultured both hESCs and hiPSCs in three different culture conditions: on mouse embryonic fibroblast (MEF) and SNL feeder cell layers together with conventional stem cell culture medium containing knockout serum replacement and basic fibroblast growth factor (bFGF), as well as on a Matrigel matrix in mTeSR1 medium. hPSC lines were subjected to cardiac differentiation in mouse visceral endodermal-like (END-2) co-cultures and the cardiac differentiation efficiency was determined by counting both the beating areas and Troponin T positive cells, as well as studying the expression of OCT-3/4, mesodermal Brachyury T and NKX2.5 and endodermal SOX-17 at various time points during END-2 differentiation by q-RT-PCR analysis. The most efficient cardiac differentiation was observed with hPSCs cultured on MEF or SNL feeder cell layers in stem cell culture medium and the least efficient cardiac differentiation was observed on a Matrigel matrix in mTeSR1 medium. Further, hPSCs cultured on a Matrigel matrix in mTeSR1 medium were found to be more committed to neural lineage than hPSCs cultured on MEF or SNL feeder cell layers. In conclusion, culture conditions have a major impact on the propensity of the hPSCs to differentiate into a cardiac lineage.Marisa OjalaKristiina RajalaMari Pekkanen-MattilaMarinka MiettinenHeini HuhtalaKatriina Aalto-SetäläPublic Library of Science (PLoS)articleMedicineRScienceQENPLoS ONE, Vol 7, Iss 10, p e48659 (2012)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Marisa Ojala Kristiina Rajala Mari Pekkanen-Mattila Marinka Miettinen Heini Huhtala Katriina Aalto-Setälä Culture conditions affect cardiac differentiation potential of human pluripotent stem cells.
description	Human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and human induced pluripotent stem cells (hiPSCs), are capable of differentiating into any cell type in the human body and thus can be used in studies of early human development, as cell models for different diseases and eventually also in regenerative medicine applications. Since the first derivation of hESCs in 1998, a variety of culture conditions have been described for the undifferentiated growth of hPSCs. In this study, we cultured both hESCs and hiPSCs in three different culture conditions: on mouse embryonic fibroblast (MEF) and SNL feeder cell layers together with conventional stem cell culture medium containing knockout serum replacement and basic fibroblast growth factor (bFGF), as well as on a Matrigel matrix in mTeSR1 medium. hPSC lines were subjected to cardiac differentiation in mouse visceral endodermal-like (END-2) co-cultures and the cardiac differentiation efficiency was determined by counting both the beating areas and Troponin T positive cells, as well as studying the expression of OCT-3/4, mesodermal Brachyury T and NKX2.5 and endodermal SOX-17 at various time points during END-2 differentiation by q-RT-PCR analysis. The most efficient cardiac differentiation was observed with hPSCs cultured on MEF or SNL feeder cell layers in stem cell culture medium and the least efficient cardiac differentiation was observed on a Matrigel matrix in mTeSR1 medium. Further, hPSCs cultured on a Matrigel matrix in mTeSR1 medium were found to be more committed to neural lineage than hPSCs cultured on MEF or SNL feeder cell layers. In conclusion, culture conditions have a major impact on the propensity of the hPSCs to differentiate into a cardiac lineage.
format	article
author	Marisa Ojala Kristiina Rajala Mari Pekkanen-Mattila Marinka Miettinen Heini Huhtala Katriina Aalto-Setälä
author_facet	Marisa Ojala Kristiina Rajala Mari Pekkanen-Mattila Marinka Miettinen Heini Huhtala Katriina Aalto-Setälä
author_sort	Marisa Ojala
title	Culture conditions affect cardiac differentiation potential of human pluripotent stem cells.
title_short	Culture conditions affect cardiac differentiation potential of human pluripotent stem cells.
title_full	Culture conditions affect cardiac differentiation potential of human pluripotent stem cells.
title_fullStr	Culture conditions affect cardiac differentiation potential of human pluripotent stem cells.
title_full_unstemmed	Culture conditions affect cardiac differentiation potential of human pluripotent stem cells.
title_sort	culture conditions affect cardiac differentiation potential of human pluripotent stem cells.
publisher	Public Library of Science (PLoS)
publishDate	2012
url	https://doaj.org/article/6b05ac8c438c4d21b5ac8017de523eb0
work_keys_str_mv	AT marisaojala cultureconditionsaffectcardiacdifferentiationpotentialofhumanpluripotentstemcells AT kristiinarajala cultureconditionsaffectcardiacdifferentiationpotentialofhumanpluripotentstemcells AT maripekkanenmattila cultureconditionsaffectcardiacdifferentiationpotentialofhumanpluripotentstemcells AT marinkamiettinen cultureconditionsaffectcardiacdifferentiationpotentialofhumanpluripotentstemcells AT heinihuhtala cultureconditionsaffectcardiacdifferentiationpotentialofhumanpluripotentstemcells AT katriinaaaltosetala cultureconditionsaffectcardiacdifferentiationpotentialofhumanpluripotentstemcells
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Culture conditions affect cardiac differentiation potential of human pluripotent stem cells.

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