Robust cardiomyocyte differentiation from human pluripotent stem cells via temporal modulation of canonical Wnt signaling

Human pluripotent stem cells (hPSCs) offer the potential to generate large numbers of functional cardiomyocytes from clonal and patient-specific cell sources. Here we show that temporal modulation of Wnt signaling is both essential and sufficient for efficient cardiac induction in hPSCs under define...

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Bibliographic Details
Published in:Proceedings of the National Academy of Sciences - PNAS 2012-07, Vol.109 (27), p.E1848-E1857
Main Authors: Lian, Xiaojun, Hsiao, Cheston, Wilson, Gisela, Zhu, Kexian, Hazeltine, Laurie B, Azarin, Samira M, Raval, Kunil K, Zhang, Jianhua, Kamp, Timothy J, Palecek, Sean P
Format: Article
Language:English
Subjects:
beta Catenin - genetics
Biological Sciences
Cardiomyocytes
Cell Culture Techniques - methods
Cell Differentiation - physiology
chemical inhibitors
Cloning
Culture Media - pharmacology
Enzyme Inhibitors - pharmacology
Glycogen Synthase Kinase 3 - antagonists & inhibitors
Glycogen Synthase Kinase 3 - metabolism
Humans
Kinases
Myocytes, Cardiac - cytology
Pluripotent Stem Cells - cytology
Pluripotent Stem Cells - metabolism
PNAS Plus
Ribonucleic acid
RNA
RNA, Small Interfering - genetics
Stem cells
tau-protein kinase
Transforming Growth Factor beta - metabolism
Wnt Signaling Pathway - drug effects
Wnt Signaling Pathway - physiology
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Summary:Human pluripotent stem cells (hPSCs) offer the potential to generate large numbers of functional cardiomyocytes from clonal and patient-specific cell sources. Here we show that temporal modulation of Wnt signaling is both essential and sufficient for efficient cardiac induction in hPSCs under defined, growth factor-free conditions. shRNA knockdown of β-catenin during the initial stage of hPSC differentiation fully blocked cardiomyocyte specification, whereas glycogen synthase kinase 3 inhibition at this point enhanced cardiomyocyte generation. Furthermore, sequential treatment of hPSCs with glycogen synthase kinase 3 inhibitors followed by inducible expression of β-catenin shRNA or chemical inhibitors of Wnt signaling produced a high yield of virtually (up to 98%) pure functional human cardiomyocytes from multiple hPSC lines. The robust ability to generate functional cardiomyocytes under defined, growth factor-free conditions solely by genetic or chemically mediated manipulation of a single developmental pathway should facilitate scalable production of cardiac cells suitable for research and regenerative applications.
ISSN:0027-8424
1091-6490
DOI:10.1073/pnas.1200250109