Induction of Human iPSC-Derived Cardiomyocyte Proliferation Revealed by Combinatorial Screening in High Density Microbioreactor Arrays

• Published in: Scientific reports Vol. 6; no. 1; p. 24637
• Main Authors: Titmarsh, Drew M., Glass, Nick R., Mills, Richard J., Hidalgo, Alejandro, Wolvetang, Ernst J., Porrello, Enzo R., Hudson, James E., Cooper-White, Justin J.
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group UK 21.04.2016; Nature Publishing Group
• Subjects: 13/100; 13/21; 14/105; 14/19; 14/56; 631/154/1435/2163; 631/61/490; Cardiomyocytes; Cell culture; Cell cycle; Cell Differentiation; Cell Proliferation; Cells, Cultured; Drug Discovery - instrumentation; Drug Discovery - methods; Fibroblast growth factors; Heart; Heart diseases; High-Throughput Screening Assays - methods; High-Throughput Screening Assays - standards; Humanities and Social Sciences; Humans; Induced Pluripotent Stem Cells - cytology; Insulin-like growth factors; Microfluidics; multidisciplinary; Myocytes, Cardiac - cytology; Myocytes, Cardiac - drug effects; Myocytes, Cardiac - metabolism; Pluripotency; Reproducibility of Results; Science; Science (multidisciplinary); Signal transduction; Signal Transduction - drug effects; Stem cell transplantation; Stem cells; Wnt protein
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/srep24637

Inducing cardiomyocyte proliferation in post-mitotic adult heart tissue is attracting significant attention as a therapeutic strategy to regenerate the heart after injury. Model animal screens have identified several candidate signalling pathways, however, it remains unclear as to what extent these pathways can be exploited, either individually or in combination, in the human system. The advent of human cardiac cells from directed differentiation of human pluripotent stem cells (hPSCs) now provides the ability to interrogate human cardiac biology in vitro , but it remains difficult with existing culture formats to simply and rapidly elucidate signalling pathway penetrance and interplay. To facilitate high-throughput combinatorial screening of candidate biologicals or factors driving relevant molecular pathways, we developed a high-density microbioreactor array (HDMA) – a microfluidic cell culture array containing 8100 culture chambers. We used HDMAs to combinatorially screen Wnt, Hedgehog, IGF and FGF pathway agonists. The Wnt activator CHIR99021 was identified as the most potent molecular inducer of human cardiomyocyte proliferation, inducing cell cycle activity marked by Ki67, and an increase in cardiomyocyte numbers compared to controls. The combination of human cardiomyocytes with the HDMA provides a versatile and rapid tool for stratifying combinations of factors for heart regeneration.