Ocular Surface Center, Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, 6565 Fannin Street, NC-205, Houston, TX 77030, USA;
Tissue engineering holds great promise for corneal transplantation to treat the blinding corneal diseases. There is a pressing need for advances in corneal tissue engineering due to an increasing shortage of donor cornea tissue. This study revealed novel strategies to isolate corneal epithelial stem cells based on their signature markers and signaling pathways, and developed a new generation of bioengineered corneal equivalents that simulate native human cornea with enriched stem cells. The isolated cell populations displayed undifferentiated phenotype, high colony forming efficiency and greater proliferative capacity with BrdU-label retaining slow-cycling stem cells. These stem/progenitor cells grew well on the donor corneal stromal lamella, prepared by FDA approved Horizon Epikeratome system. The corneal stroma was found to promote progenitor cell expansion and differentiation. An optically transparent and stratified epithelium was rapidly generated on the stromal substrate and displayed native-like morphology and structure with basal cells expressing stem/progenitor cell markers ABCG2 and p63. This corneal constructs were survival well after xeno-transplanted in the nude mouse back. In conclusion, these findings demonstrate that a novel corneal constructs have been bioengineered using isolated limbal stem cells and natural corneal stromal lamella, which simulate native human cornea and are suitable for corneal reconstruction.