HOME HELP FEEDBACK SUBSCRIPTIONS ARCHIVE SEARCH TABLE OF CONTENTS		QUICK SEARCH:   [advanced] Author: Keyword(s): Year:  Vol:  Page:

This Article Full Text Full Text (PDF) Submit a response Alert me when this article is cited Alert me when eLetters are posted Alert me if a correction is posted Citation Map Services Email this article to a friend Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Enzmann, V. Articles by Kaplan, H. J. Search for Related Content PubMed PubMed Citation Articles by Enzmann, V. Articles by Kaplan, H. J.

Enhanced Induction of RPE Lineage Markers in Pluripotent Neural Stem Cells Engrafted into the Adult Rat Subretinal Space

¹From the Department of Ophthalmology and Visual Sciences and the ²Kentucky Spinal Cord Injury Research Center, Department of Neurological Surgery, University of Louisville, Louisville, Kentucky.

PURPOSE. To investigate the differentiation of rat neural stem cells (rNSCs) into cells of retinal pigment epithelial (RPE) lineage both in vitro and in vivo, after subretinal transplantation into normal rats and in a sodium iodate (NaIO₃) model of RPE loss.

METHODS. rNSCs prepared from the cortex of embryonic day (E)14 Fisher F344 rats were cocultured with different concentrations of vasoactive intestinal peptide (VIP), adult rat RPE cells, or neurosensory retina (NSR) for 5 days. Cell morphology and expression of RPE-specific markers (cytokeratin, CD68, microphthalmia-inducing transcription factor [MITF]) were studied. Additional antibodies used to characterize the rNSCs were markers for stem cells (nestin), immature neurons (ßIII-tubulin), astrocytes (glial fibrillary acidic protein [GFAP]), and oligodendrocytes (Rip). In in vivo studies, 10⁶ green fluorescent protein [GFP]–labeled rNSCs were injected subretinally in either normal adult Lewis rats or NaIO₃-treated rats (70 mg/mL NaIO₃ administered intravenously 7 days before transplantation).

RESULTS. In vitro VIP-treated rNSCs changed from round cells to glia-like cells with processes that stained for both GFAP and nestin. In addition, small clusters of flattened, polygonal cells with an epithelial-cell–like shape that stained for cytokeratin and CD68 were observed. Coculture of rNSCs with RPE cells, but not with NSR, also led to cells of this phenotype. After transplantation, nestin⁺ and GFP⁺ rNSCs were visible subretinally as a transplant. In addition, more than 50% of transplanted rNSCs were cytokeratin⁺ and CD68⁺.

CONCLUSIONS. Very few rNSCs differentiate in vitro into epithelial-like cells that express RPE-specific markers. In vivo, this differentiation is remarkably enhanced after subretinal engraftment. Thus, transplantation of NSCs into the subretinal space may be a therapy for retinal diseases involving an RPE abnormality.

This article has been cited by other articles:

				T. Chan-Ling, L. Baxter, A. Afzal, N. Sengupta, S. Caballero, E. Rosinova, and M. B. Grant Hematopoietic Stem Cells Provide Repair Functions after Laser-Induced Bruch's Membrane Rupture Model of Choroidal Neovascularization Am. J. Pathol., March 1, 2006; 168(3): 1031 - 1044. [Abstract] [Full Text] [PDF]