Original Article

In vivo and in vitro proof for intravitreal implanted bone marrow mesenchimal stem cell (MSC) mediated gene therapy in rabbit tapetoretinal degeneration model: green fluorescent protein(GFP) - Original Article

  • Gökhan Özge
  • Riza Güngör
  • Güngör Sobaci
  • Ferit Avcu
  • Murat Demiriz
  • Elvin Akdag
  • Ali Ugur Ural

Received Date: 12.01.2008 Accepted Date: 02.07.2008 Turk J Ophthalmol 2008;38(4):320-329

Purpose: We have shown the efficacy of intravitreal use of bone marrow mesenchimal stem cell (MSC) in rabbit tapetoretinal degeneration model. In this study, the feasibility of MSC mediated gene therapy is investigated. Material-Methods: In GATA Research Center, MSCs were differantiated in vitro from bone marrow which was obtained from proximal femur of a New Zelland albino rabbit, and the cells from the forth passage were used. In cultere environment, plasmid mediated green fluorescent protein (GFP) transmission to these cells was performed two times in 48 hours. Intravenous 40 mg/kg NAIO(3) injection was done to 3 adult pigmented rabbits.(study group). Afterwards 2.5x10(5) cell/0,1 ml to right eyes and 0,1 ml balanced salt solution to left eyes intravitrealy, were performed in the study and the control eyes (1 pigmented rabbit, two eyes). Clinical, angiographic (FA-HRA 2), ERG and hystopathological evaluations were performed during the observation period (1., 5., 10., 45th day). Findings: On first day, abnormal ERG b-wave amplitudes and cluster of GFP loading cells in the vitreous were observed. On the fifth day, retinal edema, diminished b-wave amplitudes and hypofluorescent and hyperfluorescent areas on FA were noted. Immunofluorescence examination of the first rabbit showed GFP loading cells in the retinal layers. In vivo and in vitro assays done on the 10th day showed GFP loading cells in the vitreous and in the retina which had angiographically increased hypo and hyperfluorescent spots. Our controlled observation showed that these cells were still in the retinal layers on the 45th day. Results: In this acute tapetoretinal degeneration model intravitreally injected allogenic stem cells transfected by plasmid enabled GFP transmission to the damaged retina. This method may be developed as a new made of gene therapy in the retinal degeneration patients who has the advantage of using otolog MSCs.

Keywords: Gene therapy, stem cell, rabbit, tapetoretinal degeneration, green fluorescent protein GFP)

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