BACKGROUND: The transplantation of the limbus is based on the theory that limbal stem cells are necessary for epithelialisation and maintenance of a clear cornea. We used a genetic analysis procedure to demonstrate the survival of transplanted limbus stem cells. PATIENT AND METHODS: A lamellary corneal graft and limbus transplantation from donor material was carried out on a patient with severe alkali burns and destroyed Bowmann's membrane to prepare the cornea for perforating keratoplasty. Subsequently, perforating corneal graft was carried out and the epithelium and endothelium of the removed cornea were analysed with a gene analytical procedure. For the cellular typing we used the SGM kit, well known in forensic analysis, that allows simultaneous detection of alleles at 11 loci. The genetic analysis was performed using an ABI PRISM 310 Genetic Analyser (Perkin Elmer Applied Biosystems) after DNA amplification by the polymerase chain reaction. RESULTS: The cornea-limbus transplant was integrated without vascularisation from the limbus under treatment with cyclosporin A. The epithelium cells of the corneal tissue gained by keratoplasty had the same genetic pattern as the primary donor in all loci, but no congruence with the recipient (except for the amelogenin marker because both the donor and the recipient were males). Therefore the epithelium cells must have originated from the transplanted stem cells of the donor. CONCLUSIONS: The identical genetic pattern found for the donor and the epithelium of the corneal-limbal transplant demonstrate the survival of the transplanted stem cells in a patient with adequate cytostatic therapy.
BACKGROUND: The transplantation of the limbus is based on the theory that limbal stem cells are necessary for epithelialisation and maintenance of a clear cornea. We used a genetic analysis procedure to demonstrate the survival of transplanted limbus stem cells. PATIENT AND METHODS: A lamellary corneal graft and limbus transplantation from donor material was carried out on a patient with severe alkali burns and destroyed Bowmann's membrane to prepare the cornea for perforating keratoplasty. Subsequently, perforating corneal graft was carried out and the epithelium and endothelium of the removed cornea were analysed with a gene analytical procedure. For the cellular typing we used the SGM kit, well known in forensic analysis, that allows simultaneous detection of alleles at 11 loci. The genetic analysis was performed using an ABI PRISM 310 Genetic Analyser (Perkin Elmer Applied Biosystems) after DNA amplification by the polymerase chain reaction. RESULTS: The cornea-limbus transplant was integrated without vascularisation from the limbus under treatment with cyclosporin A. The epithelium cells of the corneal tissue gained by keratoplasty had the same genetic pattern as the primary donor in all loci, but no congruence with the recipient (except for the amelogenin marker because both the donor and the recipient were males). Therefore the epithelium cells must have originated from the transplanted stem cells of the donor. CONCLUSIONS: The identical genetic pattern found for the donor and the epithelium of the corneal-limbal transplant demonstrate the survival of the transplanted stem cells in a patient with adequate cytostatic therapy.