April 2011
Volume 52, Issue 14
Free
ARVO Annual Meeting Abstract  |   April 2011
Generation of Human Corneal Endothelial Cell Sheets Under Serum-Free Conditions Using Reactive And Temperature-Responsive Cell Culture Carriers
Juliane Teichmann; Monika Valtink; Stefan Gramm; Mirko Nitschke; Carsten Werner; Katrin Engelmann
Author Affiliations & Notes
  • Juliane Teichmann
    Ophthalmology, Klinikum Chemnitz gGmbH, Chemnitz, Germany
    Anatomy, Medical Faculty Carl Gustav Carus, TU Dresden, Dresden, Germany
  • Monika Valtink
    Anatomy, Medical Faculty Carl Gustav Carus, TU Dresden, Dresden, Germany
  • Stefan Gramm
    Max Bergmann Center of Biomaterials, Leibniz Institute of Polymer Research Dresden, Dresden, Germany
  • Mirko Nitschke
    Max Bergmann Center of Biomaterials, Leibniz Institute of Polymer Research Dresden, Dresden, Germany
  • Carsten Werner
    Max Bergmann Center of Biomaterials, Leibniz Institute of Polymer Research Dresden, Dresden, Germany
    CRTD Center for Regenerative Therapies Dresden, Dresden, Germany
  • Katrin Engelmann
    Ophthalmology, Klinikum Chemnitz gGmbH, Chemnitz, Germany
    CRTD Center for Regenerative Therapies Dresden, Dresden, Germany
  • Footnotes
    Commercial Relationships  Juliane Teichmann, None; Monika Valtink, None; Stefan Gramm, None; Mirko Nitschke, None; Carsten Werner, None; Katrin Engelmann, None
  • Footnotes
    Support  DFG project EN 168/13-1
Investigative Ophthalmology & Visual Science April 2011, Vol.52, 6466. doi:
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      Juliane Teichmann, Monika Valtink, Stefan Gramm, Mirko Nitschke, Carsten Werner, Katrin Engelmann; Generation of Human Corneal Endothelial Cell Sheets Under Serum-Free Conditions Using Reactive And Temperature-Responsive Cell Culture Carriers. Invest. Ophthalmol. Vis. Sci. 2011;52(14):6466.

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Abstract

Purpose: : Generation of temperature-responsive cell culture carriers with tunable biochemical and mechanical properties for the fabrication of transplantable human corneal endothelial cell sheets under serum-free conditions.

Methods: : Human corneal endothelial cells (HCEC) were cultured on electron beam cross-linked thin films of poly(vinyl methyl ether) (PVME, temperature-responsive component) blended with 1 wt% or 10 wt% of the alternating copolymer of PVME and maleic anhydride (PVME-alt-MA, protein/peptide-binding component). Film thickness and degree of cross-linking were varied to tune the mechanical properties while MA units allowed for the covalent attachment of laminin/ chondroitin sulfate, collagen type IV, fibronectin, or cyclic RGD peptide (cRGD). Cell adhesion, proliferation and detachment after lowering the temperature were monitored by microscopy and were compared for HCEC cultured with and without serum-containing growth medium (immortalized population HCEC-12 and clonal cell line HCEC-B4G12, respectively).

Results: : First experiments showed that an initial coating with cRGD gave best results in both culture systems. HCEC-12 cultured in serum-supplemented medium showed very good cell adhesion, monolayer formation and cell sheet detachment upon temperature reduction on 1 wt% PVME-alt-MA containing surfaces with various thicknesses and a medium cross linking degree as well as on thicker 10 wt% PVME-alt-MA containing surfaces. In contrast, serum-free cultured HCEC-B4G12 showed good adhesion, proliferation and detachment behavior only on thicker, high cross-linked 10 wt% PVME-alt-MA containing coatings.

Conclusions: : Temperature-responsive cell culture carriers based on electron beam cross-linked MA functionalized PVME are suitable for the generation of HCEC sheets under serum-supplemented and serum-free culture conditions.

Keywords: cornea: endothelium 
© 2011, The Association for Research in Vision and Ophthalmology, Inc., all rights reserved. Permission to republish any abstract or part of an abstract in any form must be obtained in writing from the ARVO Office prior to publication.
Copyright © 2025 Association for Research in Vision and Ophthalmology.
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