[Long-term outcome of gfp gene modified human RPE xenografts into the subretinal space of rabbits].
The possibility of delivery genes to retina by using retinal pigment epithelium (RPE) transplantation was investigated. Cultured human adult RPE cells were transfected with retrovirus encoding green fluorescent protein (GFP), then GFP-expressing RPE cells were transplanted into the subretinal space of rabbits through pars plana vitrectomy under microscopy. The transplant sites were examined in vivo by ophthalmoscope for fluorescence. The animals were euthanatized at 1, 2, 3, 4, 6, 10, 11, 14, 18, 20, 23, 24, 25, 33, 54 weeks, respectively, and the retinas were studied histologically, including epi-fluorescent microscopy, confocal microscopy and transmission electron microscopy. The GFP-expressing transplant could be followed in the living retina up to 21 days by ophthalmoscope. The epi-fluorescent microscopy examination disclosed survival of the transplanted cells in 1 year with continual and significant GFP expression. It was observed the transplanted hRPE-gfp cells were not only present at transplant sites but spread over two to three quarters of retina. Generally, RPE xenografts integrated individually into host RPE in a regular cobblestones arrangement or formed a monolayer sheet between host RPE and neural retina. It was observed that intravitreal injection of FK-506 weekly improved the survival of RPE xenografts and reduced the infiltration of mononuclear microphages and lymphocytes in choroids from 1 through 14 weeks after transplantation. The results demonstrate a long-term survival of gene modified human RPE xenografts into the subretinal space of rabbits and support the feasibility of this approach for delivery genes to retina.
Central Experimental Laboratory, The First People's Hospital of Shanghai, Shanghai 200080, China. huangq@online.sh.cn
Feng
F
Wang
Ping
P
Xu
Ji-Hong
JH
Wu
Xin
X
Xia
Hao-Long
HL
Sun
Xun
X
Xu
Qian
Q
Huang
D000328:Adult; D000818:Animals; D005123:Eye / Q000473:pathology / Q000503:physiopathology; D015870:Gene Expression; D018014:Gene Transfer Techniques; D017930:Genes, Reporter; D049452:Green Fluorescent Proteins; D006801:Humans; D008164:Luminescent Proteins / Q000235:genetics* / Q000378:metabolism; D010857:Pigment Epithelium of Eye / Q000276:immunology / Q000378:metabolism / Q000637:transplantation*; D011817:Rabbits; D012160:Retina / Q000378:metabolism / Q000473:pathology; D013997:Time Factors; D014183:Transplantation, Heterologous
D008164:Luminescent Proteins; D049452:Green Fluorescent Proteins