A simple and flexible high-throughput method for the study of cardiomyocyte proliferation

Abstract

Cardiac muscle cells lack regenerative capacity in postnatal mammals. A concerted effort has been made in the field to determine regulators of cardiomyocyte proliferation and identify therapeutic strategies to induce division, with the ultimate goal of regenerating heart tissue after a myocardial infarct. We sought to optimize a high throughput screening protocol to facilitate this effort. We developed a straight-forward high throughput screen with simple readouts to identify small molecules that modulate cardiomyocyte proliferation. We identify human induced pluripotent stem cell-derived cardiomyocytes (hiCMs) as a model system for such a screen, as a very small subset of hiCMs have the potential to proliferate. The ability of hiCMs to proliferate is density-dependent, and cell density has no effect on the outcome of proliferation: cytokinesis or binucleation. Screening a compound library revealed many regulators of proliferation and cell death. We provide a comprehensive and flexible screening procedure and cellular phenotype information for each compound. We then provide an example of steps to follow after this screen is performed, using three of the identified small molecules at various concentrations, further implicating their target kinases in cardiomyocyte proliferation. This screening platform is flexible and cost-effective, opening the field of cardiovascular cell biology to laboratories without substantial funding or specialized training, thus diversifying this scientific community.