Coculture of Endothelial Cells with Human Pluripotent Stem Cell‐Derived Cardiac Progenitors Reveals a Differentiation Stage‐Specific Enhancement of Cardiomyocyte Maturation

Abstract

Cardiomyocytes (CMs) generated from human pluripotent stem cells (hPSCs) are immature in their structure and function, limiting their potential in disease modeling, drug screening, and cardiac cellular therapies. Prior studies have demonstrated that coculture of hPSC‐derived CMs with other cardiac cell types, including endothelial cells (ECs), can accelerate CM maturation. To address whether the CM differentiation stage at which ECs are introduced affects CM maturation, the authors coculture hPSC‐derived ECs with hPSC‐derived cardiac progenitor cells (CPCs) and CMs and analyze the molecular and functional attributes of maturation. ECs have a more significant effect on acceleration of maturation when cocultured with CPCs than with CMs. EC coculture with CPCs increases CM size, expression of sarcomere, and ion channel genes and proteins, the presence of intracellular membranous extensions, and chronotropic response compared to monoculture. Maturation is accelerated with an increasing EC:CPC ratio. This study demonstrates that EC incorporation at the CPC stage of CM differentiation expedites CM maturation, leading to cells that may be better suited for in vitro and in vivo applications of hPSC‐derived CMs.

Bi-directional Impacts of Heterotypic Interactions in Engineered 3D Human Cardiac Microtissues Revealed by Single-Cell RNA-Sequencing and Functional Analysis

“…In contrast, short-term co-culture (first 7 days) with CFs promoted more rapid phenotypic maturation of CMs, in terms of both microtissue calcium handling dynamics and CM gene expression. Our finding that co-culture with ECs did not improve cardiac microtissue function is consistent with previous reports showing that while CMs may be transcriptionally or structurally impacted by EC co-culture, functional parameters are not improved 49,50 . However, a recent study demonstrated that ECs can improve stem cell-derived cardiac microtissue function when they are incorporated with a stromal population (i.e.…”

Cardiomyocyte maturation: advances in knowledge and implications for regenerative medicine

“…Interestingly, some of the benefits on cardiomyocyte maturation of co-culture with endothelial cells are specific to the hPSC-CM differentiation stage at which the endothelial cells are introduced. Dunn and colleagues found that co-culture of hPSC-derived endothelial cells with hPSC-derived cardiac progenitor cells had a greater maturation effect than co-culture with more differentiated, beating hPSC-CMs, including on cardiomyocyte size, expression of genes encoding sarcomeric and ion channel proteins, and chronotropic responses to drugs 145 . Further studies are needed with other cell types, including smooth muscle cells and peripheral neurons, to identify the roles of these cells in inducing cardiomyocyte maturation.…”

Optimizing the Use of iPSC-CMs for Cardiac Regeneration in Animal Models

“…Constructs made with co-cultures of iPSC-CMs and EMT fibroblasts had better systolic and diastolic benchmarks when compared to tissues made from iPSC-CMs and other stromal cells [158]. Likewise, adding endothelial cells and/or smooth muscle cells with the fibroblasts also improves the tissues' phenotype [159,160].…”