Purpose. We have previously shown that iris pigmented epithelial (IPE) cells can be used as an alternative to retinal pigmented epithelial (RPE) cells for transplantation into the subretinal space (IOVS 34/4, 1993, #1942). This would permit autologous transplantation thus avoiding possible graft rejection. To identify unambiguously the transplanted cells, we have used a defective herpes simplex virus (HSV) expressing the bacterial lacZ gene. Methods. Iris tissue samples of Long Evans rats were dissected and IPE cells were dissociated using trypsin digestion. They were incubated for 1 hr with lacZ-encoding HSV, washed 3 times in media, and injected into the subretinal space. After up to 4 weeks, the animals were sacrificed, the eyes were fixed, reacted with X-Gal, and processed for electron microscopy. Results. Transplanted cells were identified by their blue color under the light microscope and by crystals under the electron microscope characteristic of the X-Gal reaction product. IPE cells were found in the subretinal space including some in close contact with photoreceptor cells. Conclusions. IPE cells can be labeled using a herpes virus vector prior to transplantation. This labeling is persistent and allows the analysis of the fate of transplanted cells.
|Original language||English (US)|
|Journal||Investigative Ophthalmology and Visual Science|
|State||Published - Feb 15 1996|
ASJC Scopus subject areas
- Sensory Systems
- Cellular and Molecular Neuroscience