Maturation of Human Induced Pluripotent Stem Cell-Derived Cardiomyocytes by Soluble Factors from Human Mesenchymal Stem Cells

Abstract

In this study, we proposed that the functionality or phenotype of differentiated cardiomyocytes derived from human induced pluripotent stem cells (iPSC-CMs) might be modified by co-culture with mesenchymal stem cells (MSCs), resulting in an improved therapeutic potential for failing myocardial tissues. Structural, motility, electrophysiological, and metabolic analyses revealed that iPSC-CMs co-cultured with MSCs displayed aligned myofibrils with A-, H-, and I-bands that could contract and relax quickly, indicating the promotion of differentiation and the establishment of the iPSC-CM structural framework, and showed clear gap junctions and an electric pacing of >2 Hz, indicating enhanced cell-cell interactions. In addition, soluble factors excreted by MSCs, including several cytokines and exosomes, enhanced cardiomyocyte-specific marker production, produced more energy under normal and stressed conditions, and reduced reactive oxygen species production by iPSC-CMs under stressed condition. Notably, gene ontology and pathway analysis revealed that microRNAs and proteins in the exosomes impacted the functionality and maturation of iPSC-CMs. Furthermore, cell sheets consisting of a mixture of iPSC-CMs and MSCs showed longer survival and enhanced therapeutic effects compared with those consisting of iPSC-CMs alone. This may lead to a new type of iPSC-based cardiomyogenesis therapy for patients with heart failure.

Disruption of histamine/H1R signaling pathway represses cardiac differentiation and maturation of human induced pluripotent stem cells

“…In addition to promoting CPC formation, the histamine/H 1 R signal well presented the ability to accelerate the maturation of hiPSC-CMs, as indicated by the morphological and structural properties. The proliferation capacity of cardiomyocytes derived from hiPSCs usually declines with maturation [33,58]. There was no significant difference in the proliferation capacity observed between the histamine-treated group and the control group on the 14th day, which we thought might be because the time of differentiation was not long enough.…”

“…4a), which indicates a relatively more mature level. The protein level of gap connexins is positively correlated with the level of maturation of cardiomyocytes and is also a biomarker of maturation of cardiomyocytes [33,34]. Immunostaining of anti-connexin43 (CX43) revealed a higher level of CX43 expression in histamine-treated hiPSC-CMs (Fig.…”

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

“…In rats, formation of t‐tubules begins after birth and are fully developed after a month . However, in hPSC‐derived CMs, t‐tubules are rarely seen, and this lack has been suggested to contribute to irregular Ca 2+ handling in hPSC‐derived CMs . CMs and CPCs were generated from the H9‐hTnnTZ‐pGZ‐D2, an hESC line that expresses eGFP under the TNNT2 promoter, and used in monoculture and coculture configurations with hPSC‐derived ECs as illustrated in Figure A,B.…”

“…Heterotypic intercellular interactions also impact CM maturation. Coculture of mesenchymal stem cells with induced pluripotent stem cell (iPSC)‐derived CMs increased CM contractility and sarcomere organization and alignment . This effect was recapitulated with EC‐derived exosomes containing proteins and microRNAs .…”

“…Coculture of mesenchymal stem cells with induced pluripotent stem cell (iPSC)‐derived CMs increased CM contractility and sarcomere organization and alignment . This effect was recapitulated with EC‐derived exosomes containing proteins and microRNAs . Similarly, fibroblasts have been shown to induce hPSC‐derived CM maturation, including elevated cardiac troponin T (cTnT), Cx43, and α‐actinin protein expression, and contractility through both soluble factors and extracellular matrix protein production …”