title: 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 α-5 and α-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.
authors.affiliation: Institute for Stem Cell and Regenerative Medicine, University of Washington, Seattle, WA 98109, USA.
authors.forename: Kara White
authors.initials: KW
authors.lastname: Moyes
authors.forename: Christopher G
authors.initials: CG
authors.lastname: Sip
authors.forename: Willimark
authors.initials: W
authors.lastname: Obenza
authors.forename: Emily
authors.initials: E
authors.lastname: Yang
authors.forename: Cody
authors.initials: C
authors.lastname: Horst
authors.forename: Robert E
authors.initials: RE
authors.lastname: Welikson
authors.forename: Stephen D
authors.initials: SD
authors.lastname: Hauschka
authors.forename: Albert
authors.initials: A
authors.lastname: Folch
authors.forename: Michael A
authors.initials: MA
authors.lastname: Laflamme
mesh_terms: D057134:Antibodies, Neutralizing / Q000494:pharmacology; D002448:Cell Adhesion; D002454:Cell Differentiation; D002465:Cell Movement / Q000187:drug effects; D015194:Diffusion Chambers, Culture; D053595:Embryonic Stem Cells / Q000166:cytology* / Q000378:metabolism; D005109:Extracellular Matrix / Q000187:drug effects* / Q000235:genetics / Q000378:metabolism; D005353:Fibronectins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D015870:Gene Expression; D006801:Humans; D057054:Molecular Imaging; D032383:Myocytes, Cardiac / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D017469:Receptors, Fibronectin / Q000037:antagonists & inhibitors / Q000235:genetics / Q000378:metabolism; D015398:Signal Transduction; D051153:Wnt Proteins / Q000235:genetics / Q000378:metabolism / Q000494:pharmacology*; D000071818:Wnt-5a Protein
chemical_list: D057134:Antibodies, Neutralizing; D005353:Fibronectins; D011518:Proto-Oncogene Proteins; D017469:Receptors, Fibronectin; C085389:WNT5A protein, human; D051153:Wnt Proteins; D000071818:Wnt-5a Protein