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Chi-Chun Lai, Peter Gouras, Ken Doi, Fang Lu, Hild Kjeldbye, Steven P. Goff, Robert Pawliuk, Philippe Leboulch, Stephen H. Tsang; Tracking RPE Transplants Labeled by Retroviral Gene Transfer with Green Fluorescent Protein. Invest. Ophthalmol. Vis. Sci. 1999;40(9):2141-2146.
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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
methods. Cultured human fetal retinal pigment epithelium (HFRPE) was
exposed to green fluorescent protein (GFP)-transducing retroviral
vectors, Moloney murine leukemia virus, and lentivirus. The cultured
cells were followed by fluorescence microscopy. Suspensions of
GFP-expressing HFRPE were transplanted into the subretinal space of
pigmented rabbits, and the transplant sites were examined by SLO for
fluorescence, including fluorescein and indocyanine green
angiography. The rabbits were euthanatized at different times after
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
least 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 periods of
time by retroviral gene transfer. GFP allowed retinal transplants and
gene expression to be monitored in vivo. These results provide a model
for potential ex vivo gene therapy in the subretinal
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