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V. Enzmann, R.M. Howard, Y. Yamauchi, S.R. Whittemore, H.J. Kaplan; Rat Neural Stem Cells (rNSC) Express RPE Marker After Subretinal Transplantation . Invest. Ophthalmol. Vis. Sci. 2003;44(13):502.
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© ARVO (1962-2015); The Authors (2016-present)
Purpose: To investigate the differentiation of rNSC in vivo after subretinal transplantation in normal rats and in a sodium iodate (NaIO3) model of RPE loss. Methods: rNSC prepared from the cortex of embryonic Fisher F344 rats (E14) were maintained in DMEM/F12 medium containing FGF-2 and N2 supplement. The cells were GFP-transfected using a retroviral vector. Approx. 1x106 cells (passage 1-3) in 20 µl medium were injected subretinally using a sterile glass pipette in Lewis rats (n=5 per group). NaIO3-treated Lewis rats received 70 mg/kg NaIO3 iv 7 days prior to transplantation. Light microscopy and immunohistochemistry evaluated RPE cell loss and the differentiation of transplanted cells - rNSC: nestin+ and GFP+; RPE cells: cytokeratin+, CD68+, and MITF+. Results: Treatment with NaIO3 led to disruption of RPE monolayer 3 days after injection and to significant RPE loss after 7 days. rNSC were visible subretinally on light microscopy (9/10) 7 days after transplantation and were nestin+ (9/10) and GFP+ (3/10). Variation in the transfection rate probably accounted for the increased staining for nestin. Transplanted cells were also cytokeratin+ (8/10) and CD68+ (8/10), as was the host RPE monolayer (5/5). MITF+ cells were found in the transplants, too (4/10). Conclusions: After transplantation into the subretinal space of normal and NaIO3-treated rats, rNSC express the RPE cell marker cytokeratin, CD68, and MITF. These observations suggest that NSC may replace lost RPE cells in non-exudative AMD, as well as in exudative AMD following submacular surgery.
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