Human pluripotent stem cells (hPSC) are investigated as a source of authentic human cardiac cells for drug discovery and toxicological tests. Cell-based assays using automated fluorescence imaging platform and high-content analysis characterize hypertrophic and toxicity profiles of compounds in hPSC-derived cardiomyocytes (hPSC-CM) at the cellular and subcellular levels. In purified population of hPSC-CM loaded with cell tracer probe and cell death markers, both hypertrophic and toxicity profiles can be assessed in live cardiomyocyte cultures. Alternatively, in non-purified cultures of hPSC-CM, hypertrophy, proliferation, and cell death assays can be performed specifically in the cardiomyocyte subpopulation using antibodies directed against cardiac proteins and a combination of cell death- and proliferation-specific fluorescent probes.
Human pluripotent stem cell Cardiomyocyte Hypertrophy Cell death Proliferation Automated high-content imaging
Springer Nature is developing a new tool to find and evaluate Protocols. Learn more
G.F. was supported by Fondation Leducq and Hungarian Scientific Research Fund (OTKA 105555) and TÁMOP-4.2.2/B-10/1-2010-0013 Grant.
Burridge PW, Keller G, Gold JD et al (2012) Production of de novo cardiomyocytes: human pluripotent stem cell differentiation and direct reprogramming. Cell Stem Cell 10:16–28PubMedCrossRefGoogle Scholar
Matsa E, Denning C (2012) In vitro uses of human pluripotent stem cell-derived cardiomyocytes. J Cardiovasc Transl Res 5:581–592PubMedCrossRefGoogle Scholar
Laflamme MA, Chen KY, Naumova AV et al (2007) Cardiomyocytes derived from human embryonic stem cells in pro-survival factors enhance function of infarcted rat hearts. Nat Biotechnol 25:1015–1024PubMedCrossRefGoogle Scholar
Mioulane M, Foldes G, Ali NN et al (2012) Quantification of cell death mechanisms in human pluripotent stem cell-derived cardiomyocytes. J Cardiovasc Transl Res 5:593–604PubMedCrossRefGoogle Scholar
Kehat I, Kenyagin-Karsenti D, Snir M et al (2001) Human embryonic stem cells can differentiate into myocytes with structural and functional properties of cardiomyocytes. J Clin Invest 108:407–414PubMedGoogle Scholar
Hattori F, Chen H, Yamashita H et al (2010) Nongenetic method for purifying stem cell-derived cardiomyocytes. Nat Methods 7:61–66PubMedCrossRefGoogle Scholar
Foldes G, Mioulane M, Wright JS et al (2011) Modulation of human embryonic stem cell-derived cardiomyocyte growth: a testbed for studying human cardiac hypertrophy? J Mol Cell Cardiol 50:367–376PubMedCrossRefGoogle Scholar