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S. Keijser, G.F. J. M. Vrensen, F.A. Prins, H.J. Tanke, R.J. W. Keizer de, M.J. Jager; A New Limbal Transplant Model, Using E–GFP for in vivo Monitoring of Transplant Survival . Invest. Ophthalmol. Vis. Sci. 2005;46(13):3586.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: Limbal transplants in humans show a high rate of rejection, even under local and systemic immunotherapy. In order to test experimentally immuno–modulatory treatments we developed a new limbal transplant model in the rat. Donor material was obtained from a transgenic rat, and enhanced green fluorescent protein (E–GFP) was used for in vivo follow up of the limbal transplant. Methods: Eighteen E–GFP positive limbal transplants from a Sprague–Dawley TgN(act–EGFP)Osb4 rat were transplanted on 18 Fischer 344 (allogenic). The transplant size was about 1.5 mm by 3.5 mm, and the transplant was sutured with 10.0 nylon to the limbal region. Six rats were monitored three times a week using a fluorescent microscope, equipped with a digital camera, until E–FGP fluorescence has fully disappeared. At regular intervals, rats were sacrificed for immunohistochemistry. Results: In vivo monitoring showed that during the first week the transplant size of most allogenic transplants remained stable. Hereafter the transplant size decreased and the allogenic transplants were completely rejected around post operative day 15. The immunohistochemical observations confirmed the in vivo observations. Conclusions: With a modified fluorescent microscope we were able to take digital pictures of E–GFP positive transplant, and therefore monitor the transplant behavior over time, without disturbance of the ocular surface. We successfully developed a new limbal transplant model, in which the transplant can be accurately followed in vivo, without the necessity of taking corneal material for PCR or immunohistochemistry. This model will be used to test new therapies for limbal transplant survival.
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