Abstract
Isolating actively proliferating cardioblasts is the first crucial step for cardiac regeneration through cell implantation. However, the origin and identity of putative cardioblasts are still unclear. Here, we uncover a novel class of cardiac lineage cells, PDGFRα+Flk1− cardioblasts (PCBs), from mouse and human pluripotent stem cells induced using CsAYTE, a combination of the small molecules Cyclosporin A, the rho-associated coiled-coil kinase inhibitor Y27632, the antioxidant Trolox, and the ALK5 inhibitor EW7197. This novel population of actively proliferating cells is cardiac lineage–committed but in a morphologically and functionally immature state compared to mature cardiomyocytes. Most important, most of CsAYTE-induced PCBs spontaneously differentiated into functional αMHC+ cardiomyocytes (M+CMs) and could be a potential cellular resource for cardiac regeneration.
Highlights
A, the rho-associated coiled-coil kinase inhibitor Y27632, the antioxidant Trolox, and the ALK5 inhibitor EW7197
For monitoring and tracing cardiac lineage cells (CLCs) commitment, we used EMG7 embryonic stem cells (ESCs), which have a transgene consisting of cardiac-specific αmyosin heavy chain promoter–driven enhanced green fluorescent protein (GFP); in addition, we screened for the cardiac-specific markers cardiac troponin T and α-actinin in differentiating pluripotent stem cells (PSCs)
CsAYTE could efficiently induce and expand PDGFRα+Flk1− cardioblasts (PCBs) but only for a short period, and they lost their proliferative potential after fully differentiating cardiomyocytes
Summary
A, the rho-associated coiled-coil kinase inhibitor Y27632, the antioxidant Trolox, and the ALK5 inhibitor EW7197. Cardiovascular diseases remain a leading cause of mortality worldwide, with death arising from the inability of cardiomyocytes to regenerate after myocardial injury In this aspect, cardiac lineage cells (CLCs) from pluripotent stem cells (PSCs) have become the most attractive cellular resource underlying an unprecedented strategy in cell-based therapy to rescue damaged hearts[1,2,3]. We screened for various signaling modulators and established a novel, simple, and efficient method for cardiac lineage specification from mouse PSC-derived Flk1+ MPCs using a combination of four specific modulators: CsA, the ROCK inhibitor Y27632, the antioxidant Trolox, and the activin A receptor type II-like kinase (ALK5) inhibitor EW7197 (referred to collectively here as CsAYTE). We showed that CsAYTE-induced PCBs could proliferate and could spontaneously further differentiate into functional cardiomyocytes with high efficiency, which can be a novel cellular resource for cardiac regeneration
Sign-in/Register to view highlights & summary Sign-in/Register to access full text options 
Free) 
Talk to us
Join us for a 30 min session where you can share your feedback and ask us any queries you have
Schedule a call
