Measuring hypertrophy in neonatal rat primary cardiomyocytes and human iPSC-derived cardiomyocytes

• Published in: Methods (San Diego, Calif.) Vol. 203; pp. 447 - 464
• Main Authors: Bourque, Kyla, Hawey, Cara, Jones-Tabah, Jace, Pétrin, Darlaine, Martin, Ryan D., Ling Sun, Yi, Hébert, Terence E.
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.07.2022
• Subjects: Hypertrophy; Imaging; Induced pluripotent stem cells; Primary neonatal cardiomyocytes; RNCM; MOA-A/H2O2/ERK; Nkx2-5; ESC-CM; CAMKII; ACTA; NF-κB; Oct4; Sox2; FBS; Imaging; BNP or NPPB; MYBPC3; ZAK-MKK7-ANF axis; ACTN2; ACTN1; PKA; PKC; α-MHC or MYH6; CDX-2; PPARα; SERCA; β-MHC or MYH7; IL-6; ANP or ANF or NPPA; ERK1/2; ET-1; HCM; UII; Induced pluripotent stem cells; MYL2; cTNNT; GPCR; PCR; Hypertrophy; TAGLN; c-myc; Gβ1; cAMP; EGFR; NFAT; P-TEFb; iPSCs; Eomes; TGF-β; TNNI3; MSX1; Wnt; Brd4; JunD; Primary neonatal cardiomyocytes; Gγ2; STAT3; Klf4; Ang II; PLCβ; NFATC4; GSK3β; ACTC1; TAC
• ISSN: 1046-2023; 1095-9130
• DOI: 10.1016/j.ymeth.2021.12.006

[Display omitted] •We provide protocols to isolate rat neonatal cardiomyocytes and to derive cardiomyocytes from human iPSCs.•We show how to use high content imaging to characterize hypertrophic responses.•Combining high content approaches with measures of gene expression changes can track hypertrophic responses. In the heart, left ventricular hypertrophy is initially an adaptive mechanism that increases wall thickness to preserve normal cardiac output and function in the face of coronary artery disease or hypertension. Cardiac hypertrophy develops in response to pressure and volume overload but can also be seen in inherited cardiomyopathies. As the wall thickens, it becomes stiffer impairing the distribution of oxygenated blood to the rest of the body. With complex cellular signalling and transcriptional networks involved in the establishment of the hypertrophic state, several model systems have been developed to better understand the molecular drivers of disease. Immortalized cardiomyocyte cell lines, primary rodent and larger animal models have all helped understand the pathological mechanisms underlying cardiac hypertrophy. Induced pluripotent stem cell-derived cardiomyocytes are also used and have the additional benefit of providing access to human samples with direct disease relevance as when generated from patients suffering from hypertrophic cardiomyopathies. Here, we briefly review in vitro and in vivo model systems that have been used to model hypertrophy and provide detailed methods to isolate primary neonatal rat cardiomyocytes as well as to generate cardiomyocytes from human iPSCs. We also describe how to model hypertrophy in a “dish” using gene expression analysis and immunofluorescence combined with automated high-content imaging.