title: In vitro expansion of corneal endothelial cells on biomimetic substrates.
abstract: Corneal endothelial (CE) cells do not divide in vivo, leading to edema, corneal clouding and vision loss when the density drops below a critical level. The endothelium can be replaced by transplanting allogeneic tissue; however, access to donated tissue is limited worldwide resulting in critical need for new sources of corneal grafts. In vitro expansion of CE cells is a potential solution, but is challenging due to limited proliferation and loss of phenotype in vitro via endothelial to mesenchymal transformation (EMT) and senescence. We hypothesized that a bioengineered substrate recapitulating chemo-mechanical properties of Descemet's membrane would improve the in vitro expansion of CE cells while maintaining phenotype. Results show that bovine CE cells cultured on a polydimethylsiloxane surface with elastic modulus of 50 kPa and collagen IV coating achieved >3000-fold expansion. Cells grew in higher-density monolayers with polygonal morphology and ZO-1 localization at cell-cell junctions in contrast to control cells on polystyrene that lost these phenotypic markers coupled with increased α-smooth muscle actin expression and fibronectin fibril assembly. In total, these results demonstrate that a biomimetic substrate presenting native basement membrane ECM proteins and mechanical environment may be a key element in bioengineering functional CE layers for potential therapeutic applications.
authors.affiliation: 1] Department of Biomedical Engineering, Carnegie Mellon University, Pittsburgh, PA 15213 [2] Department of Ophthalmology, University of Pittsburgh, Pittsburgh PA 15213 [3] Louis J. Fox Center for Vision Restoration, Pittsburgh PA 15213.
authors.forename: Rachelle N
authors.initials: RN
authors.lastname: Palchesko
authors.affiliation: 1] Department of Ophthalmology, University of Pittsburgh, Pittsburgh PA 15213 [2] Louis J. Fox Center for Vision Restoration, Pittsburgh PA 15213.
authors.forename: Kira L
authors.initials: KL
authors.lastname: Lathrop
authors.affiliation: 1] Department of Ophthalmology, University of Pittsburgh, Pittsburgh PA 15213 [2] Louis J. Fox Center for Vision Restoration, Pittsburgh PA 15213.
authors.forename: James L
authors.initials: JL
authors.lastname: Funderburgh
authors.affiliation: 1] Department of Biomedical Engineering, Carnegie Mellon University, Pittsburgh, PA 15213 [2] Department of Materials Science and Engineering, Carnegie Mellon University, Pittsburgh PA 15213.
authors.forename: Adam W
authors.initials: AW
authors.lastname: Feinberg
mesh_terms: D000199:Actins / Q000096:biosynthesis; D000818:Animals; D032701:Biomimetics; D002417:Cattle; D018929:Cell Culture Techniques; D003315:Cornea / Q000473:pathology*; D016039:Corneal Transplantation; D004129:Dimethylpolysiloxanes / Q000737:chemistry; D042783:Endothelial Cells / Q000166:cytology / Q000637:transplantation*; D005353:Fibronectins / Q000096:biosynthesis; D006801:Humans; D014184:Transplantation, Homologous
chemical_list: D000199:Actins; D004129:Dimethylpolysiloxanes; D005353:Fibronectins; C013830:baysilon