Human Embryonic Stem Cell-Derived Cardiomyocytes Migrate in Response to Gradients of Fibronectin and Wnt5a

Abstract

An improved understanding of the factors that regulate the migration of human embryonic stem cell-derived cardiomyocytes (hESC-CMs) would provide new insights into human heart development and suggest novel strategies to improve their electromechanical integration after intracardiac transplantation. Since nothing has been reported as to the factors controlling hESC-CM migration, we hypothesized that hESC-CMs would migrate in response to the extracellular matrix and soluble signaling molecules previously implicated in heart morphogenesis. To test this, we screened candidate factors by transwell assay for effects on hESC-CM motility, followed by validation via live-cell imaging and/or gap-closure assays. Fibronectin (FN) elicited a haptotactic response from hESC-CMs, with cells seeded on a steep FN gradient showing nearly a fivefold greater migratory activity than cells on uniform FN. Studies with neutralizing antibodies indicated that adhesion and migration on FN are mediated by integrins a-5 and a-V. Next, we screened 10 soluble candidate factors by transwell assay and found that the noncanonical Wnt, Wnt5a, elicited an approximately twofold increase in migration over controls. This effect was confirmed using the gap-closure assay, in which Wnt5a-treated hESC-CMs showed approximately twofold greater closure than untreated cells. Studies with microfluidic-generated Wnt5a gradients showed that this factor was chemoattractive as well as chemokinetic, and Wnt5a-mediated responses were inhibited by the Frizzled-1/2 receptor antagonist, UM206. In summary, hESC-CMs show robust promigratory responses to FN and Wnt5a, findings that have implications on both cardiac development and cell-based therapies.

Anti-Cancer Phytometabolites Targeting Cancer Stem Cells

“…Additionally, CD44 + CD24 - CSCs from head and neck cancer exhibit overexpression of WNT5A protein; while WNT5A is knocking-down CSCs population was reduced [119, 120]. Recently, it was observed that Rspo2, an endogenous ligand, and 6-bromoindirubin-3'-oxime (BIO), a GSK3β inhibitor, produced a pronounced effect on pluripotency markers in lung adenocarcinoma cells (SPC-A1 and PC-9).…”

Integrins and Integrin-Associated Proteins in the Cardiac Myocyte

“…could greatly affect the reported expression patterns and this must be considered. Additional work has shown that hES-derived CMs migrate in response to fibronectin and both integrin α5 and αv 161 .…”

Mesenchymal Stem Cells and Their Conditioned Medium Improve Integration of Purified Induced Pluripotent Stem Cell–Derived Cardiomyocyte Clusters into Myocardial Tissue

“…For example, stromal-cell-derived factor 1 has been found to increase after myocardial infarction and in heart failure [37][38][39], and its chemoattractant properties have been demonstrated to enhance the recruitment of circulating hematopoietic stem cells to the infarcted myocardium [40,41]. In a recent study, Wnt5a, which is expressed also by MSCs [41,42], was identified as a chemoattractant for human ES-CMs [43]. The addition of MSCs, which migrate and integrate into the myocardial tissue, might therefore help to activate a migration of transplanted CMs, explaining at least in part the superior integration of iPS-CMs into nonvital slice tissue found in the present study.…”