Transplantation into the subretinal space of iris pigmented epithelial cells labeled using a lacZ-encoding defective HSV

M. Gelanze, P. Meneses, M. Brittis, M. R. Rosenfeld, Robert Duvoisin, D. J. Coleman

Research output: Contribution to journalArticle

Abstract

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 languageEnglish (US)
JournalInvestigative Ophthalmology and Visual Science
Volume37
Issue number3
StatePublished - Feb 15 1996
Externally publishedYes

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Defective Viruses
Iris
Simplexvirus
Transplantation
Epithelial Cells
Bacterial Genes
Long Evans Rats
Photoreceptor Cells
Lac Operon
Autologous Transplantation
Cell Transplantation
Graft Rejection
Trypsin
Digestion
Electron Microscopy
Color
Electrons
Viruses
Light

ASJC Scopus subject areas

  • Ophthalmology

Cite this

Transplantation into the subretinal space of iris pigmented epithelial cells labeled using a lacZ-encoding defective HSV. / Gelanze, M.; Meneses, P.; Brittis, M.; Rosenfeld, M. R.; Duvoisin, Robert; Coleman, D. J.

In: Investigative Ophthalmology and Visual Science, Vol. 37, No. 3, 15.02.1996.

Research output: Contribution to journalArticle

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AU - Meneses, P.

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AU - Rosenfeld, M. R.

AU - Duvoisin, Robert

AU - Coleman, D. J.

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