IOVS Molecular Human Reproduction
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(Investigative Ophthalmology and Visual Science. 2006;47:1646-1652.)
© 2006 by The Association for Research in Vision and Ophthalmology, Inc.
DOI:  10.1167/iovs.05-1092
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Retinal Pigment Epithelium Damage Enhances Expression of Chemoattractants and Migration of Bone Marrow–Derived Stem Cells

Yang Li,1 Ryan G. Reca,2 Pelin Atmaca-Sonmez,1 Mariusz Z. Ratajczak,2 Suzanne T. Ildstad,3 Henry J. Kaplan,1 and Volker Enzmann1

1From the Department of Ophthalmology and Visual Sciences, the 2Stem Cell Program, Department of Medicine, and the 3Institute for Cellular Therapeutics, University of Louisville, Louisville, Kentucky.

PURPOSE. To characterize chemoattractants expressed by the retinal pigment epithelium (RPE) after sodium iodate (NaIO3)–induced damage and to investigate whether ocular-committed stem cells preexist in the bone marrow (BM) and migrate in response to the chemoattractive signals expressed by the damaged RPE.

METHODS. C57/BL6 mice were treated with a single intravenous injection of NaIO3 (50 mg/kg) to create RPE damage. At different time points real-time RT-PCR, ELISA, and immunohistochemistry were used to identify chemoattractants secreted in the subretinal space. Conditioned medium from NaIO3-treated mouse RPE was used in an in vitro assay to assess chemotaxis of stem cell antigen-1 positive (Sca-1+) BM mononuclear cells (MNCs). The expression of early ocular markers (MITF, Pax-6, Six-3, Otx) in migrated cells and in MNCs isolated from granulocyte colony–stimulating factor (G-CSF) and Flt3 ligand (FL)–mobilized and nonmobilized peripheral blood (PB) was analyzed by real-time RT-PCR.

RESULTS. mRNA for stromal cell–derived factor-1 (SDF-1), C3, hepatocyte growth factor (HGF), and leukemia inhibitory factor (LIF) was significantly increased, and higher SDF-1 and C3 protein secretion from the RPE was found after NaIO3treatment. A higher number of BMMNCs expressing early ocular markers migrated to conditioned medium from damaged retina. There was also increased expression of early ocular markers in PBMNCs after mobilization.

CONCLUSIONS. Damaged RPE secretes cytokines that have been shown to serve as chemoattractants for BM-derived stem cells (BMSCs). Retina-committed stem cells appear to reside in the BM and can be mobilized into the PB by G-CSF and FL. These stem cells may have the potential to serve as an endogenous source for tissue regeneration after RPE damage.




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