title: Transforming growth factor β and platelet-derived growth factor modulation of myofibroblast development from corneal fibroblasts in vitro.
abstract: The purpose of this study was to test the hypotheses that development of mature vimentin+/α-smooth muscle actin+/desmin+ (V+A+D+) myofibroblasts from corneal fibroblasts is regulated by transforming growth factor (TGF) β and platelet-derived growth factor (PDGF); and that myofibroblast development in vitro follows a similar developmental pathway as it does in vivo. Mouse corneal stromal fibroblasts (MSF) were isolated from the corneas of Swiss Webster mice and cultured in serum-free media augmented with DMEM/F12 and varying doses of TGFβ (0.1-2.0 ng/ml), with and without mouse PDGF-AA and/or PDGF-BB (2.0 ng/ml), to study the transition of the MSF to V+A+D+ myofibroblasts. The mean percentage of vimentin+, α-SMA+ and desmin+ cells was determined at each time point (2-15 days), with each growth factor concentration. MSF in vitro were noted to undergo the same developmental transition from V+A-D- to V+A+D- to V+A+D+ myofibroblasts as precursors undergo in vivo. TGFβ at a dose of 0.5 ng/ml and 1.0 ng/ml with 2.0 ng/ml PDGF-AA and 2.0 ng/ml PDGF-BB in DMEM/F12 serum-free media was optimal for the development of V+A+D+ myofibroblasts. This study defines optimal in vitro conditions to monitor the development of MSF into myofibroblasts. The combined effects of TGFβ and PDGF promote the full development of V+A+D+ myofibroblasts from MSF.
authors.affiliation: Cole Eye Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
authors.forename: Vivek
authors.initials: V
authors.lastname: Singh
authors.affiliation: Cole Eye Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
authors.forename: Flavia L
authors.initials: FL
authors.lastname: Barbosa
authors.affiliation: Cole Eye Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
authors.forename: Andre A M
authors.initials: AA
authors.lastname: Torricelli
authors.affiliation: Cole Eye Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
authors.forename: Marcony R
authors.initials: MR
authors.lastname: Santhiago
authors.affiliation: Cole Eye Institute, Cleveland Clinic, Cleveland, OH 44195, USA. Electronic address: wilsons4@ccf.org.
authors.forename: Steven E
authors.initials: SE
authors.lastname: Wilson
mesh_terms: D000199:Actins / Q000378:metabolism; D000818:Animals; D000077214:Becaplermin; D002454:Cell Differentiation / Q000187:drug effects*; D002478:Cells, Cultured; D060527:Corneal Keratocytes / Q000166:cytology* / Q000378:metabolism; D003319:Corneal Stroma / Q000166:cytology; D003893:Desmin / Q000378:metabolism; D004338:Drug Combinations; D007150:Immunohistochemistry; D051379:Mice; D058628:Myofibroblasts / Q000166:cytology* / Q000378:metabolism; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology*; D020574:Proto-Oncogene Proteins c-sis / Q000494:pharmacology*; D016212:Transforming Growth Factor beta / Q000494:pharmacology*; D014746:Vimentin / Q000378:metabolism
chemical_list: D000199:Actins; D003893:Desmin; D004338:Drug Combinations; D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; D016212:Transforming Growth Factor beta; D014746:Vimentin; C544013:alpha-smooth muscle actin, mouse; C075278:platelet-derived growth factor A; D000077214:Becaplermin
keywords: corneal stroma; corneal wound healing; development; myofibroblasts; platelet-derived growth factor; transforming growth factor beta