Tracking RPE transplants labeled by retroviral gene transfer with green fluorescent protein

PURPOSE. TO determine whether human retinal pigment epithelium (RPE) can be modified by retroviral-mediated gene transfer and to monitor the human RPE cells in the subretinal space of living rabbits with scanning laser ophtha lmoscopy (SLO). METHODS. Cultured human fetal retinal pigment epithelium (HFRPE) was expose d to green fluorescent protein (GFP)-transducing retroviral vectors, Molone y murine leukemia virus, and lentivirus. The cultured cells were followed b y fluorescence microscopy. Suspensions of GFP-expressing HFRPE were transpl anted into the subretinal space of pigmented rabbits, and the transplant si tes were examined by SLO for fluorescence, including fluorescein and indocy anine green angiography. The rabbits were euthanatized at different times a fter transplantation, and the retinas were studied histologically. RESULTS. Retroviral gene transfer can introduce a foreign gene such as GFP into cultured HFRPE. Gene expression is maintained in cultured RPE for at l east 3 months. The lentiviral vector traduced both nondividing and dividing cells; the Moloney vector only transduced the latter. GFP-expressing cells can be followed in the living retina. Their changes reflect the rejection response followed histologically. CONCLUSIONS. Cultured HFRPE could be transduced to express GFP for long per iods of time by retroviral gene transfer. GFP allowed retinal transplants a nd gene expression to be monitored in vivo. These results provide a model f or potential ex vivo gene therapy in the subretinal space.