May 2003
Volume 44, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2003
Rat Neural Stem Cells (rNSC) Express RPE Marker After Subretinal Transplantation
Author Affiliations & Notes
  • V. Enzmann
    Ophthalmology, University of Louisville, Louisville, KY, United States
  • R.M. Howard
    Neurological Surgery, University of Louisville, Louisville, KY, United States
  • Y. Yamauchi
    Neurological Surgery, University of Louisville, Louisville, KY, United States
  • S.R. Whittemore
    Neurological Surgery, University of Louisville, Louisville, KY, United States
  • H.J. Kaplan
    Neurological Surgery, University of Louisville, Louisville, KY, United States
  • Footnotes
    Commercial Relationships  V. Enzmann, None; R.M. Howard, None; Y. Yamauchi, None; S.R. Whittemore, None; H.J. Kaplan, None.
  • Footnotes
    Support  Research to Prevent Blindness; Commonwealth of Kentucky Research Challenge Fund
Investigative Ophthalmology & Visual Science May 2003, Vol.44, 502. doi:
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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)

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Abstract

Abstract: : 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.

Keywords: retinal degenerations: cell biology • animal model • transplantation 
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