Stage-Specific Optimization of Activin/Nodal and BMP Signaling Promotes Cardiac Differentiation of Mouse and Human Pluripotent Stem Cell Lines

Abstract

Efficient differentiation of embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs) to a variety of lineages requires step-wise approaches replicating the key commitment stages found during embryonic development. Here we show that expression of PdgfR-a segregates mouse ESC-derived Flk-1 mesoderm into Flk-1 + PdgfR-a + cardiac and Flk-1 + PdgfR-a À hematopoietic subpopulations. By monitoring Flk-1 and PdgfR-a expression, we found that specification of cardiac mesoderm and cardiomyocytes is determined by remarkably small changes in levels of Activin/Nodal and BMP signaling. Translation to human ESCs and iPSCs revealed that the emergence of cardiac mesoderm could also be monitored by coexpression of KDR and PDGFR-a and that this process was similarly dependent on optimal levels of Activin/Nodal and BMP signaling. Importantly, we found that individual mouse and human pluripotent stem cell lines require optimization of these signaling pathways for efficient cardiac differentiation, illustrating a principle that may well apply in other contexts.

Eomesodermin induces Mesp1 expression and cardiac differentiation from embryonic stem cells in the absence of Activin

“…Altogether, the three types of cells derived from multipotent cardiovascular progenitors (MCPs) [27,[35][36][37] were massively increased following Eomes expression in DDM, although we cannot rule out that some endothelial and smooth muscle cells could also derive from other progenitors (such as the hemangioblasts [38] that could also be induced following Eomes expression). We and others have recently demonstrated that a combination of monoclonal antibodies (Flk1/Pdgfra) can be used to isolate the earliest Mesp1 expressing MCPs arising during ESC differentiation [16,27]. FACS analysis of ESC 48 h after Eomes overexpression in DDM revealed a significant enrichment for Flk1/ Pdgfra-positive MCPs in Eomes overexpressing cells (Fig 1O, supplementary Fig S2E online), consistent with the increase of Mesp1 expressing MCPs by Eomes in these conditions.…”

Global DNA methylation and transcriptional analyses of human ESC-derived cardiomyocytes

“…Recent efforts have successfully developed several protocols, by which hESCs can be differentiated into hCMs (Yang et al, 2008;Kattman et al, 2011;Lian et al, 2012;Willems et al, 2012;Zhang et al, 2012). Furthermore, hESC-derived hCMs have been analyzed by microarray and a group of cardiospecific genes were revealed (Beqqali et al, 2006;Cao et al, 2008;Synnergren et al, 2008).…”

Future Treatment of Heart Failure Using Human iPSC-Derived Cardiomyocytes

“…However, this chemical alone might be not enough to induce cardiac differentiation from iPSCs. Several studies have shown that various combinations of heart developmentrelated proteins including BMP, activin, Wnt, BMP inhibitor, and Wnt inhibitor induce cardiomyocytes from ESCs [7][8][9][10]. We reported that the context-dependent differential action of BMPs in cardiomyocyte induction is explained by the local action of Noggin and other BMP inhibitors and, accordingly, developed a protocol to induce cardiac differentiation of mouse ESCs through transient administration of Noggin [9].…”