Purchase this article with an account.
M. Notara, N. A. Bullett, P. Deshpande, D. B. Haddow, S. MacNeil, J. T. Daniels; Plasma Polymer-Coated Contact Lenses for Serum-Free Culture and Transplantation of Limbal Epithelium. Invest. Ophthalmol. Vis. Sci. 2007;48(13):1886.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
The aim of this work is to develop a plasma polymer-coated contact lens system for the serum-free culture and transplantation of limbal epithelial stem cells (LESC). This novel serum-free approach provides a culture surface as well as a cell-transfer vehicle.
A range of plasma polymer surfaces including acrylic acid, allyl alcohol, allyl amine and octa-1, 7-diene were fabricated at various monomer flow rates and power and their surface chemistry was examined using XPS analysis. A human corneal epithelial cell line (HCE-C) was used as a preliminary screening tool in order to identify the optimum surface type. The MTT-ESTA assay was used to assess proliferation. Primary human corneal epithelial cells (HCE) were isolated from cadaveric tissue provided by Moorfields Eye bank and co-cultured with growth arrested 3T3 mouse fibroblasts. Immunocytochemistry methods were used for primary cell characterization. A rabbit in vitro organ culture (IVOC) model was used to assess the transfer of cultured HCE cells from the plasma-coated contact lenses to a corneal wound bed.
MTT_ESTA data suggested superior proliferation rates of HCE-C on the acrylic acid surfaces. HCE cells exhibited good cell confluence and proliferation on acrylic acid plasma coated surfaces in the presence of serum. Under serum-free conditions, the tissue-culture plastic substrate was inferior to all the acrylic acid coated surfaces, in terms of cell confluence and metabolic activity as assessed by MTT-ESTA assay. The HCE cells cultured on all substrates expressed high levels of ABCG2 and delta-NP63-alpha and low levels of cytokeratin 3 on all the surfaces studied suggesting low levels of differentiation.HCE cells were able to grow and proliferate on the entire surface of acrylic acid coated contact lenses forming confluent epithelial sheets. Cell transfer has been successful on the IVOC wound beds for subconfluent cultures.
The above data suggest that this novel LESC transplantation system merits further investigation as a promising alternative for cornea reconstitution surgery.
This PDF is available to Subscribers Only