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K.–H. Chen; Transplantation of Human Corneal Endothelial Cells on a Thermo–Responsive Surface . Invest. Ophthalmol. Vis. Sci. 2005;46(13):4528.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: o investigate the feasibility for harvesting human corneal endothelial (HCE) cell sheet with intact cell junctions via temperature modulation of thermo–responsive polymer–grafted surface. Methods: Apoly–N–isopropylacrylamide (PNIPAAm)–grafted polyethylene (PE) surface was prepared by plasma induced graft polymerization. The optimal conditions for surface modification were characterized by peroxide destruction studies; static water contact angle measurements; and functional group analyses. HCE cells from donor cornea were isolated and cultured on the PNIPAAm–graft PE surface at 37°C. After reaching confluence, the detachment of HCE cell sheet from thermo–responsive surface was attributed to the lowering temperature effect. The viability and tight junction of detached cell sheet were assessed by Live/Dead Viability/Cytotoxicity kits and zonula occludins–1 (ZO–1) immuno–staining reagents, respectively Results: The confluent human corneal endothelial ccells could be harvested as a whole sheet after the temperature was decreased to 21°C. The detached cell sheet was shown a nornmal architecture of a nomolayer and good cell viability. Conclusions: The technology for harvesting HCE cell sheet via temperature modulation of thermo–responsive surface was successfully developed.
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