Efficient differentiation of cardiomyocytes and generation of calcium-sensor reporter lines from nonhuman primate iPSCs

Nonhuman primate (NHP) models are more predictive than rodent models for developing induced pluripotent stem cell (iPSC)-based cell therapy, but robust and reproducible NHP iPSC-cardiomyocyte differentiation protocols are lacking for cardiomyopathies research. We developed a method to differentiate...

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Bibliographic Details
Published in:Scientific reports 2018-04, Vol.8 (1), p.5907-16, Article 5907
Main Authors: Lin, Yongshun, Liu, Huimin, Klein, Michael, Ostrominski, John, Hong, So Gun, Yada, Ravi Chandra, Chen, Guibin, Navarengom, Keron, Schwartzbeck, Robin, San, Hong, Yu, Zu-Xi, Liu, Chengyu, Linask, Kaari, Beers, Jeanette, Qiu, Lugui, Dunbar, Cynthia E., Boehm, Manfred, Zou, Jizhong
Format: Article
Language:English
Subjects:
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631/532/1360
631/532/2064/2158
Animal models
Calcium (intracellular)
Cardiomyocytes
CRISPR
Drug screening
Heart diseases
Humanities and Social Sciences
multidisciplinary
Myocardial infarction
Pluripotency
Science
Science (multidisciplinary)
Stem cells
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Summary:Nonhuman primate (NHP) models are more predictive than rodent models for developing induced pluripotent stem cell (iPSC)-based cell therapy, but robust and reproducible NHP iPSC-cardiomyocyte differentiation protocols are lacking for cardiomyopathies research. We developed a method to differentiate integration-free rhesus macaque iPSCs (RhiPSCs) into cardiomyocytes with >85% purity in 10 days, using fully chemically defined conditions. To enable visualization of intracellular calcium flux in beating cardiomyocytes, we used CRISPR/Cas9 to stably knock-in genetically encoded calcium indicators at the rhesus AAVS1 safe harbor locus. Rhesus cardiomyocytes derived by our stepwise differentiation method express signature cardiac markers and show normal electrochemical coupling. They are responsive to cardiorelevant drugs and can be successfully engrafted in a mouse myocardial infarction model. Our approach provides a powerful tool for generation of NHP iPSC-derived cardiomyocytes amenable to utilization in basic research and preclinical studies, including in vivo tissue regeneration models and drug screening.
ISSN:2045-2322
2045-2322
DOI:10.1038/s41598-018-24074-y