Generation of PDGFRα+ Cardioblasts from Pluripotent Stem Cells

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.

Trolox‐induced cardiac differentiation is mediated by the inhibition of Wnt/β‐catenin signaling in human embryonic stem cells

“…While previous studies have shown that a single dose of trolox induces low cardiac differentiation efficiency in FACS‐sorted mesodermal cells derived from murine and human PSCs (Cho et al, ; Hong et al, ), our results showed much higher cardiac differentiation efficiency following a single dose of trolox. As the differentiation conditions or cell manipulation processes employed in the present study are different from those in previous studies, the difference in the cardiac differentiation efficiency could be related to the methodological difference.…”

Regenerative potential of mouse embryonic stem cell-derived PDGFRα+ cardiac lineage committed cells in infarcted myocardium

“…Flk1 + MPCs were sorted by AutoMACS Pro Separator (Miltenyi Biotec). For induction of CLCs, sorted Flk1 + MPCs were plated onto the mitomycin C (AG Scientific)-treated confluent OP9 cells at a density of 5-10 × 10 3 cells cm 2 in the medium containing 3 μg/mL of CsA, 10 μmol/L of Y27632, 400 μmol/L of Trolox, and 1 μg/mL of EW7197 (CsAYTE)[11,17]. The medium was refreshed every other day.…”

“…Recently, several studies have been conducted to identify a novel marker for cardiac progenitor or cardiac lineage-committed cells (CLCs), which are intermediate-stage cells between mesodermal cells and differentiated CMs with proliferative capacity[7-10]. Our group previously established a novel class of cells from PSCs-platelet-derived growth factor receptor-α (PDGFRα) + CLCs-induced using a combination of four specific modulators: the mitochondrial permeability transition pore inhibitor cyclosporin A (CsA), the ROCK inhibitor Y27632, the antioxidant Trolox, and the activin A receptor type II-like kinase (ALK5) inhibitor EW7197 (collectively referred to here as CsAYTE)[11]. This novel population of actively proliferating cells is cardiac lineage-committed but in a morphologically and functionally immature state compared with differentiated CMs[11].…”

“…Our group previously established a novel class of cells from PSCs-platelet-derived growth factor receptor-α (PDGFRα) + CLCs-induced using a combination of four specific modulators: the mitochondrial permeability transition pore inhibitor cyclosporin A (CsA), the ROCK inhibitor Y27632, the antioxidant Trolox, and the activin A receptor type II-like kinase (ALK5) inhibitor EW7197 (collectively referred to here as CsAYTE)[11]. This novel population of actively proliferating cells is cardiac lineage-committed but in a morphologically and functionally immature state compared with differentiated CMs[11]. In the present study, we investigated the regenerative potential of mouse ESC-derived PDGFRα + CLCs in a murine MI model and compared their efficacy with differentiated CMs.…”

A Novel Atypical PKC-Iota Inhibitor, Echinochrome A, Enhances Cardiomyocyte Differentiation from Mouse Embryonic Stem Cells

“…Pluripotent stem cells (PSCs), including embryonic stem cells and induced pluripotent stem cells, are attractive cellular sources for cardiac stem cell therapy [ 5 ]. Various chemical compounds have been studied as potential regulators of the differentiation of PSCs into cardiac lineages, and most of these chemical compounds have been related to diverse signaling pathways involving the bone morphogenetic protein, transforming growth factor, activin, nodal, Wnt, rho-associated coiled-coil kinase, and fibroblast growth factor [ 5 , 6 ]. However, cardiogenic molecules extracted from natural sources remain to be investigated.…”