Hille K, Makuch D, Wilske J, Ruprecht K W
Universitäts-Augenklinik, Kirrberger Strasse 1, 66421 Homburg/Saar.
Ophthalmologe. 2002 Jul;99(7):575-9. doi: 10.1007/s003470100548.
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.
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.
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.
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.
角膜缘移植基于这样一种理论,即角膜缘干细胞对于角膜上皮化和维持透明角膜是必需的。我们使用基因分析程序来证明移植的角膜缘干细胞的存活情况。
对一名患有严重碱烧伤且Bowman膜受损的患者进行了板层角膜移植和取自供体材料的角膜缘移植,为穿透性角膜移植术准备角膜。随后进行了穿透性角膜移植,并使用基因分析程序对切除角膜的上皮和内皮进行分析。对于细胞分型,我们使用了法医分析中熟知的SGM试剂盒,该试剂盒可同时检测11个位点的等位基因。在通过聚合酶链反应进行DNA扩增后,使用ABI PRISM 310基因分析仪(珀金埃尔默应用生物系统公司)进行基因分析。
在用环孢素A治疗的情况下,角膜-角膜缘移植在没有角膜缘血管化的情况下实现了整合。角膜移植术后获得的角膜组织的上皮细胞在所有位点上具有与原供体相同的基因模式,但与受体不一致(除牙釉蛋白标记物外,因为供体和受体均为男性)。因此,上皮细胞必定起源于供体移植的干细胞。
在接受充分细胞抑制治疗的患者中,供体与角膜-角膜缘移植上皮细胞的相同基因模式证明了移植干细胞的存活。