Chemically Defined Production of Tri‐Lineage Human iPSC‐Derived Cardiac Spheroids

Abstract

AbstractCardiac spheroids derived from human induced pluripotent stem cells (hiPSC‐cardiac spheroids) represent a powerful three‐dimensional (3D) model for examining cardiac physiology and for drug toxicity screening. Recent advances with self‐organizing, multicellular cardiac organoids highlight the capability of directed stem cell differentiation approaches to recapitulate the composition of the human heart in vitro. Using hiPSC‐derived cardiomyocytes (hiPSC‐CMs), hiPSC‐derived endothelial cells (hiPSC‐ECs), and hiPSC‐derived cardiac fibroblasts (hiPSC‐CFs) is advantageous for enabling tri‐cellular crosstalk within a multilineage system and for generating patient‐specific models. Chemically defined medium containing factors needed to simultaneously maintain hiPSC‐CMs, hiPSC‐ECs, and hiPSC‐CFs is used to produce the spheroid system. In this article, we present protocols to illustrate the methods for conducting small‐molecule‐mediated differentiations of hiPSCs into cardiomyocytes, endothelial cells, and cardiac fibroblasts, as well as to assemble the fully integrated cardiac spheroids. © 2023 Wiley Periodicals LLC.Basic Protocol 1: Maintenance and expansion of hiPSCsBasic Protocol 2: Differentiation of hiPSCs into cardiomyocytesBasic Protocol 3: Differentiation of hiPSCs into vascular endothelial cellsBasic Protocol 4: Differentiation of hiPSCs into cardiac fibroblastsBasic Protocol 5: Production of hiPSC‐derived cardiac spheroids