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V. Enzmann, P. Atmaca–Sonmez, Y. Li, T.H. Shehan, C. Schanie, S.T. Ildstad, H.J. Kaplan; In vitro and in vivo Differentiation of Bone Marrow–Derived Stem Cells Into Retinal Pigment Epithelial–Like Cells . Invest. Ophthalmol. Vis. Sci. 2006;47(13):1769.
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To determine if bone marrow–derived stem cells (BMSC) have the capacity in vitro and in vivo to express retinal pigment epithelial (RPE)–like markers.
In vitro, mouse Sca–1+ GFP+ cells of bone marrow origin were used in coculture with adult mouse RPE cells. The coculture in a 1:1 ratio was performed with and without cell–cell–contact for up to 3 weeks. Mouse fibroblasts served as a control. Immunocytochemical analysis was performed using monoclonal antibodies (mAbs) against specific RPE markers – cytokeratin, RPE65, MITF – as well as non–RPE markers – opsin (photoreceptors) and glial fibrillary acidic protein (GFAP; glia). In vivo, sodium iodate (NaIO3) was used to damage the RPE. For this study, C57BL/6 mice were injected i.v. with 35 mg/kg NaIO3 followed by the subretinal (s.r.) injection of 3x104 Sca–1+ GFP+ BMSC on day 3. The mice were sacrificed on days 7, 14, 21, and 28 after transplantation. Whole eye flat mounts (FM) were prepared and examined for GFP+ cells under a laser scanning microscope (argon laser). Immunocytochemical analysis was performed on FM, as well as on paraffin cross sections, using mAbs against the specific RPE marker RPE65.
In vitro, BMSC changed from round to flattened, polygonal cells and expressed cytokeratin, RPE65 and MITF when cocultured in direct contact with RPE cells. In vivo, with 35 mg/ml of NaIO3 patchy RPE damage was observed starting on day 14. At the same time point the transplanted GFP+ Sca–1+ BMSC in the subretinal space at the sites of RPE loss expressed RPE65.
BMSC are capable of expressing RPE–like markers in coculture with adult RPE cells, as well as in the subretinal space in a murine model of RPE cell loss. However, it is not yet known whether these cells are capable of replacing the damaged RPE functionally.
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