Journal article
Long-term studies on allotransplantation of rabbit retinal pigment epithelial cells double-labelled with 5-bromodeoxyuridine and natural pigment
Chinese medical journal, Vol.111(8), pp.736-740
01 Aug 1998
PMID: 11245031
Featured in Collection : UN Sustainable Development Goals @ Drexel
Abstract
OBJECTIVE
To facilitate the identification of transplanted retinal pigment epithelial (RPE) cells, we sought to double-label the cells with 5-bromodeoxyuridine (BrdU) and with natural pigment. The BrdU is not lost during cell division but does require immunohistochemical methods for visualization; the pigment on the other hand, allows immediate, obvious identification, but is gradually lost with cell division. Together they provide a convenient, long-term double label.Non-confluent RPE cells at the second to the fifth passages were labelled with 5-BrdU and pigment. The double-labelled RPE cells were transplanted onto Bruch's membrane of 72 eyes of New Zealand albino rabbits. The labelled cells were localized by anti-BrdU antibody and the avidin biotin-alkaline phosphatase complex (ABC-AP) method, and by visible inclusions of pigment.
METHODS
Non-confluent RPE cells at the second to the fifth passages were labelled with 5-BrdU and pigment. The double-labelled RPE cells were transplanted onto Bruch's membrane of 72 eyes of New Zealand albino rabbits. The labelled cells were localized by anti-BrdU antibody and the avidin biotin-alkaline phosphatase complex (ABC-AP) method, and by visible inclusions of pigment.The transplanted RPE cells had distinct basal and apical morphology, and were in close contact with the photoreceptor outer segments of the host. The BrdU label was restricted to the nuclei of the RPE cells, which were stained blue. The pigment was located in the cytoplasm of the apical portion of these RPE cells. No evidence of severe rejection was seen.
RESULTS
The transplanted RPE cells had distinct basal and apical morphology, and were in close contact with the photoreceptor outer segments of the host. The BrdU label was restricted to the nuclei of the RPE cells, which were stained blue. The pigment was located in the cytoplasm of the apical portion of these RPE cells. No evidence of severe rejection was seen.Using this double-label method, transplanted RPE cells could be readily and reliably identified till one year after transplantation. The transplanted RPE cells revealed normal morphology with some function: they had distinct basal and apical morphology as seen in close contact with the outer segment of photoreceptor cells. The junctional complexes were well formed with neighboring RPE cells. The transplanted RPE cells phagocytose shed outer segments of the host. No evidence of rejection was observed, suggesting that the subretinal space of the rabbit may possess some degree of immunologic privilege. This experiment provides reliable evidence for the clinical research of allotransplantation of RPE cells.
CONCLUSIONS
Using this double-label method, transplanted RPE cells could be readily and reliably identified till one year after transplantation. The transplanted RPE cells revealed normal morphology with some function: they had distinct basal and apical morphology as seen in close contact with the outer segment of photoreceptor cells. The junctional complexes were well formed with neighboring RPE cells. The transplanted RPE cells phagocytose shed outer segments of the host. No evidence of rejection was observed, suggesting that the subretinal space of the rabbit may possess some degree of immunologic privilege. This experiment provides reliable evidence for the clinical research of allotransplantation of RPE cells.
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Details
- Title
- Long-term studies on allotransplantation of rabbit retinal pigment epithelial cells double-labelled with 5-bromodeoxyuridine and natural pigment
- Creators
- J Ye - Peking Union Medical College HospitalW Li - Hahnemann University HospitalS J Ryan - University of Southern California
- Publication Details
- Chinese medical journal, Vol.111(8), pp.736-740
- Number of pages
- 5
- Resource Type
- Journal article
- Language
- English
- Academic Unit
- Ophthalmology [Historical]
- Web of Science ID
- WOS:000075504700016
- Scopus ID
- 2-s2.0-0032129564
- Other Identifier
- 991019299107704721
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- Collaboration types
- Domestic collaboration
- International collaboration
- Web of Science research areas
- Ophthalmology