Abstract
Purpose:
To delineate the best technique for culturing and carrying human corneal limbal epithelial cells and induced pluripotent stem (iPS) cells on contact lenses (CLs) for the purpose of cell transplantation for limbal stem cell deficiency (LSCD).
Methods:
Limbal explant sections from corneoscleral rims, remaining from corneal transplantation, or induced pluripotent stem (iPS) cells were placed on the inner aspect of 5 types of CLs coated with or without 0.1% gelatin. The CLs used included lotrafilcon A (Air Optix Night & Day® Aqua), vifilcon A (Focus Monthly®), lotrafilcon A (Focus Night & Day® Aqua), etafilcon A (1-Day Acuvue®) and nelfilcon A (Focus Dailies®). Plastic culture plates and the human amniotic membrane were used, respectively, as growth platform controls for the CLs. Comparison between the growth patterns on the different CLs and the gelatin coating was made with regard to epithelial proliferation rate and epithelial cell morphology utilizing light and electron microscopy.
Results:
All the growth platforms yield monolayers culture cells with uniform epithelial cell morphology. There were marked differences in growth patterns between the different CLs. The explants placed on silicone hydrogel CL lotrafilcon A (CIBA Vision Corporation, Duluth, GA, USA) showed the fastest proliferative and migratory rates, with CLs covered with gelatin. Explants applied on lotrafilcon A were tightly attached to it, and growth was detected first on day 4. Lotrafilcon A (Air Optix Night & Day® Aqua) reached 91.2±8.5% confluence by day 16±2, and Lotrafilcon A (Focus Night & Day®) reached 93.7±6.2% confluence by day 14±3. iPS cells which were seeded on CLs were attached to lotrafilcon A by day 3. Cells examined under light microscopy were morphologically similar to corneal epithelium grown on 6 wells culture plate and amniotic membrane. Electron microscopy revealed cell-to-cell links and multiple cell layers. Microvilli and cell projections were also evident and most probably served as anchorage points on the CLs surface.
Conclusions:
Silicone hydrogel CL lotrafilcon A was advantageous over the other types of CLs for culturing and carrying corneal limbal epithelial and iPS cells. These CLs may be used as possible carriers for cultured human ocular surface epithelial cell sheets as a part of a cell therapy strategy in LSCD.
Keywords: 477 contact lens •
482 cornea: epithelium •
721 stem cells