Experimental rat model for therapeutic retinal pigment epithelium transplantation--unequivocal microscopic identification of human donor cells by in situ hybridisation of human-specific Alu sequences.
Transplantation of retinal pigment epithelial (RPE) cells is discussed as a possible therapeutic approach for retinal degeneration. Xenogeneic transplantation of human RPE cells in animal models has been studied extensively. Various methods have been used to identify the graft cells, but these methods interfere with cell behaviour so that the monitored physiological post-transplantation course may be influenced. In the present study, we applied a method for an unequivocal identification of the graft cells without interfering cell metabolism or behaviour using in situ hybridisation (ISH) of human specific Alu sequences. Visualisation of the strong extended nuclear signal of Alu sequences was much easier than that of the small nuclear signals of donor-specific sex chromosome probes. With Alu probe, even single graft cells can be identified and their development can be observed in short-term and long-term studies. With this procedure, we could prove that donor cells were injected correctly into the subretinal space by a special injection technique that we developed previously. In combination with immunohistochemistry, donor cells could be clearly discriminated from macrophages, which contained phagocytosed donor cell fragments. Application of these ISH methods for species-specific identification was valuable for follow-up-studies of RPE transplantation.
Warncke
Beate
B
Department of Pathology, University Hospital Hamburg-Eppendorf, Martinistrasse 52, 20246 Hamburg, Germany.
Valtink
Monika
M
Weichel
Judith
J
Engelmann
Katrin
K
Schäfer
Hansjörg
H
D020087:Alu Elements; D000818:Animals; D000911:Antibodies, Monoclonal; D002467:Cell Nucleus / Q000737:chemistry; D004247:DNA / Q000032:analysis; D006801:Humans; D007150:Immunohistochemistry; D017403:In Situ Hybridization; D008264:Macrophages / Q000276:immunology; D023421:Models, Animal; D010587:Phagocytosis; D010857:Pigment Epithelium of Eye / Q000637:transplantation* / Q000648:ultrastructure; D051381:Rats; D014019:Tissue Donors; D014183:Transplantation, Heterologous
D000911:Antibodies, Monoclonal; D004247:DNA