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T. Kawakita, S. Shimmura, K. Higa, J. Shimazaki, K. Tsubota; The Fate of Trans-Amplifying Cells Analyzed by in vivo Rabbit Model With Stainless Ring Transplantation. Invest. Ophthalmol. Vis. Sci. 2008;49(13):6064. doi: https://doi.org/.
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© ARVO (1962-2015); The Authors (2016-present)
To investigate the fate of trans-amplifying epithelial cells in the central cornea using a rabbit persistent corneal epithelial defect model created by transplanting a stainless steel ring in the cornea. Using this model, epithelial cells outside the ring failed to migrate inside the ring.
A stainless steel ring (8 mm diameter, 300µm wide and 250µm depth) was transplanted into rabbit corneal stroma using 10-0 nylon interrupted sutures. Subsequently we observed wound healing of epithelial defects created within the ring (4, 5, 6, 8mm diameter) compared with control corneas without ring transplants. After 1 week, hematoxylin staining and phenotype analysis was performed by immunostaining for p63, Ki67, and CK3 in a set of animals. Others were continuously observed with repeated epithelial defect of same diameter until cells exhausted.
Corneal stroma with ring transplantation for 1week showed little inflammation. When epithelium was totally removed inside of the ring, the epithelial defect persisted for at least a month. (n=3) Immunostaining showed similar expression of p63, Ki67, and CK3 compared to normal control. Wound healing was observed in smaller defects (4, 5, 6 mm), however, once cells failed to cover the surface after repeated epithelial removal, cells exhausted on the next day in all cases.
Trans-amplifying cells may be maintained for more than 1 month when separated from limbal stem cells in vivo. Their fate may depend on replicative senescence, as well as the area required to heal.
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