title: Thermo-responsive cell culture carriers based on poly(vinyl methyl ether)-the effect of biomolecular ligands to balance cell adhesion and stimulated detachment.
abstract: Two established material systems for thermally stimulated detachment of adherent cells were combined in a cross-linked polymer blend to merge favorable properties. Through this approach poly(N-isopropylacrylamide) (PNiPAAm) with its superior switching characteristic was paired with a poly(vinyl methyl ether)-based composition that allows adjusting physico-chemical and biomolecular properties in a wide range. Beyond pure PNiPAAm, the proposed thermo-responsive coating provides thickness, stiffness and swelling behavior, as well as an apposite density of reactive sites for biomolecular functionalization, as effective tuning parameters to meet specific requirements of a particular cell type regarding initial adhesion and ease of detachment. To illustrate the strength of this approach, the novel cell culture carrier was applied to generate transplantable sheets of human corneal endothelial cells (HCEC). Sheets were grown, detached, and transferred onto planar targets. Cell morphology, viability and functionality were analyzed by immunocytochemistry and determination of transepithelial electrical resistance (TEER) before and after sheet detachment and transfer. HCEC layers showed regular morphology with appropriate TEER. Cells were positive for function-associated marker proteins ZO-1, Na+/K+-ATPase, and paxillin, and extracellular matrix proteins fibronectin, laminin and collagen type IV before and after transfer. Sheet detachment and transfer did not impair cell viability. Subsequently, a potential application in ophthalmology was demonstrated by transplantation onto de-endothelialized porcine corneasin vitro. The novel thermo-responsive cell culture carrier facilitates the generation and transfer of functional HCEC sheets. This paves the way to generate tissue engineered human corneal endothelium as an alternative transplant source for endothelial keratoplasty.
authors.affiliation: Institute of Anatomy, Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden, Fetscherstraße 74, D-01307 Dresden, Germany; Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials, Hohe Straße 6, D-01069 Dresden, Germany.
authors.forename: Juliane
authors.initials: J
authors.lastname: Teichmann
authors.affiliation: Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials, Hohe Straße 6, D-01069 Dresden, Germany.
authors.forename: Mirko
authors.initials: M
authors.lastname: Nitschke
authors.affiliation: Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials, Hohe Straße 6, D-01069 Dresden, Germany.
authors.forename: Dagmar
authors.initials: D
authors.lastname: Pette
authors.affiliation: Institute of Anatomy, Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden, Fetscherstraße 74, D-01307 Dresden, Germany.
authors.forename: Monika
authors.initials: M
authors.lastname: Valtink
authors.affiliation: Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials, Hohe Straße 6, D-01069 Dresden, Germany; Herlac Coswig GmbH, Industriestraße 28, D-01640 Coswig, Germany.
authors.forename: Stefan
authors.initials: S
authors.lastname: Gramm
authors.affiliation: Institute of Physiology, Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden, Fetscherstraße 74, D-01307 Dresden, Germany.
authors.forename: Frauke V
authors.initials: FV
authors.lastname: Härtel
authors.affiliation: Institute of Physiology, Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden, Fetscherstraße 74, D-01307 Dresden, Germany.
authors.forename: Thomas
authors.initials: T
authors.lastname: Noll
authors.affiliation: Institute of Anatomy, Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden, Fetscherstraße 74, D-01307 Dresden, Germany; CRTD/DFG-Center for Regenerative Therapies Dresden-Cluster of Excellence, Fetscherstraße 105, D-01307 Dresden, Germany.
authors.forename: Richard H W
authors.initials: RH
authors.lastname: Funk
authors.affiliation: Institute of Anatomy, Faculty of Medicine Carl Gustav Carus, Technische Universität Dresden, Fetscherstraße 74, D-01307 Dresden, Germany; CRTD/DFG-Center for Regenerative Therapies Dresden-Cluster of Excellence, Fetscherstraße 105, D-01307 Dresden, Germany; Department of Ophthalmology, Klinikum Chemnitz gGmbH, Flemmingstraße 2, D-09116 Chemnitz, Germany.
authors.forename: Katrin
authors.initials: K
authors.lastname: Engelmann
authors.affiliation: Leibniz Institute of Polymer Research Dresden, Max Bergmann Center of Biomaterials, Hohe Straße 6, D-01069 Dresden, Germany; CRTD/DFG-Center for Regenerative Therapies Dresden-Cluster of Excellence, Fetscherstraße 105, D-01307 Dresden, Germany.
authors.forename: Carsten
authors.initials: C
authors.lastname: Werner
keywords: biomolecular functionalization; corneal endothelial cell sheet; endothelial transplantation; physico-chemical properties; poly(N-isopropylacrylamide); thermo-responsive polymer; tissue engineering