title: Nanofiber-based in vitro system for high myogenic differentiation of human embryonic stem cells.
abstract: Myogenic progenitor cells derived from human embryonic stem cells (hESCs) can provide unlimited sources of cells in muscle regeneration but their clinical uses are largely hindered by the lack of efficient methods to induce differentiation of stem cells into myogenic cells. We present a novel approach to effectively enhance myogenic differentiation of human embryonic stem cells using aligned chitosan-polycaprolactone (C-PCL) nanofibers constructed to resemble the microenvironment of the native muscle extracellular matrix (ECM) in concert with Wnt3a protein. The myogenic differentiation was assessed by cell morphology, gene activities, and protein expression. hESCs grown on C-PCL uniaxially aligned nanofibers in media containing Wnt3a displayed an elongated morphology uniformly aligned in the direction of fiber orientation, with increased expressions of marker genes and proteins associated with myogenic differentiation as compared to control substrates. The combination of Wnt3a signaling and aligned C-PCL nanofibers resulted in high percentages of myogenic-protein expressing cells over total treated hESCs (83% My5, 91% Myf6, 83% myogenin, and 63% MHC) after 2 days of cell culture. Significantly, this unprecedented high-level and fast myogenic differentiation of hESC was demonstrated in a culture medium containing no feeder cells. This study suggests that chitosan-based aligned nanofibers combined with Wnt3a can potentially act as a model system for embryonic myogenesis and muscle regeneration.
authors.affiliation: Department of Materials Science & Engineering, University of Washington , Seattle, Washington 98195, United States.
authors.forename: Matthew
authors.initials: M
authors.lastname: Leung
authors.forename: Ashleigh
authors.initials: A
authors.lastname: Cooper
authors.forename: Soumen
authors.initials: S
authors.lastname: Jana
authors.forename: Ching-Ting
authors.initials: CT
authors.lastname: Tsao
authors.forename: Timothy A
authors.initials: TA
authors.lastname: Petrie
authors.forename: Miqin
authors.initials: M
authors.lastname: Zhang
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation; D049109:Cell Proliferation; D048430:Cell Shape; D002478:Cells, Cultured; D053595:Embryonic Stem Cells / Q000502:physiology*; D006801:Humans; D017570:MyoD Protein / Q000378:metabolism; D032448:Myoblasts, Skeletal / Q000378:metabolism*; D057139:Nanofibers / Q000737:chemistry*; D059467:Transcriptome; D060449:Wnt Signaling Pathway; D060509:Wnt3A Protein / Q000378:metabolism
chemical_list: D017570:MyoD Protein; C057363:MyoD1 myogenic differentiation protein; C555510:WNT3A protein, human; D060509:Wnt3A Protein