Advertisement

PEDF Promotes Retinal Neurosphere Formation and Expansion In Vitro

  • Anna De Marzo
  • Claudia Aruta
  • Valeria Marigo
Chapter
Part of the Advances in Experimental Medicine and Biology book series (AEMB, volume 664)

Abstract

The retina is subject to degenerative conditions leading to blindness. Although retinal regeneration is possible in lower vertebrates, it does not occur in the adult mammalian retina. Retinal stem cell (RSC) research offers unique opportunities for developing clinical application for therapy. The ciliary body of adult mammals represents a source of quiescent RSC. These neural progenitors have a limited self-renewal potential in vitro but this can be improved by mitogens. Pigment Epithelium Derived Factor (PEDF), a member of the serpin gene family, is synthesized and secreted by retinal pigment epithelium (RPE) cells. We tested combinations of PEDF with fibroblast growth factor (FGF) during RSC growth to evaluate self-renewal and subsequent differentiation into retinal-like neuronal cell types. Medium supplemented with FGF + PEDF enhanced the RSC yield and more interestingly allowed expansion of the culture by increasing secondary retinal neurospheres after the 1st passage. This effect was accompanied by cell proliferation as revealed by BrdU incorporation. PEDF usage did not affect rod-like differentiation potential. This was demonstrated by immunofluorescence analysis of Rhodopsin and Pde6b that were found similarly expressed in cells derived from FGF or FGF + PEDF cultured RSC. Our studies suggest a possible application of PEDF in Retinal Stem Cell culture and transplantation.

Keywords

Fibroblast Growth Factor Neural Stem Cell Ciliary Body Pigment Epithelium Derive Factor Neurosphere Formation 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

Notes

Acknowledgments

We are grateful to Fondazione Cassa di Risparmio di Modena for Dr De Marzo fellowship support and XIII International Symposium on Retinal Degeneration for her travel award grant. We acknowledge the CIGS of University of Modena for providing confocal microscopy assistance. This work was supported by research grants EVI-GENORET: LSHG-CT-2005-512036 from the European Community, by research grant GGP06096 from Fondazione Telethon and PRIN 2006053302_003.

References

  1. Ahmad I, Tang L, Pham H (2000) Identification of neural progenitors in the adult mammalian eye. Biochem Biophys Res Commun 270:517–521CrossRefPubMedGoogle Scholar
  2. Arsenijevic Y (2003) Mammalian neural stem-cell renewal: nature versus nurture. Mol Neurobiol 27(1):73–98CrossRefPubMedGoogle Scholar
  3. Becerra SP, Sagasti A, Spinella P et al (1995) Pigment epithelium-derived factor behaves like a noninhibitory serpin. Neurotrophic activity does not require the serpin reactive loop. J Biol Chem 270:25992–25999PubMedGoogle Scholar
  4. Coles BL, Angénieux B, Inoue T, Del Rio-Tsonis K, Spence JR, McInnes RR, Arsenijevic Y, van der Kooy D (2004) Facile isolation and the characterization of human retinal stem cells. Proc Natl Acad Sci U S A 101(44):15772–15777CrossRefPubMedGoogle Scholar
  5. Giordano F, De Marzo A, Vetrini F et al (2007) Facile isolation and the characterization of human retinal stem cells. Mol Vis 13:1842–1850PubMedGoogle Scholar
  6. Gu P, Harwood LG, Zhang X et al (2007) Isolation of retinal progenitor and stem cells from the porcine eye. Mol Vis 13:1045–1057PubMedGoogle Scholar
  7. Inoue Y, Yanagi Y, Tamaki Y et al (2005) Clonogenic analysis of ciliary epithelial derived retinal progenitor cells in rabbits. Exp Eye Res 81(4):437–445CrossRefPubMedGoogle Scholar
  8. Kokkinopoulos I, Pearson RA, Macneil A, Dhomen NS, Maclaren RE, Ali RR, Sowden JC (2008) Isolation and characterisation of neural progenitor cells from the adult Chx10(orJ/orJ) central neural retina. Mol Cell Neurosci 38(3):359–373Google Scholar
  9. Livne-Bar I, Pacal M, Cheung MC et al (2006) Chx10 is required to block photoreceptor differentiation but is dispensable for progenitor proliferation in the postnatal retina. Proc Natl Acad Sci U S A 103(13):4988–4993CrossRefPubMedGoogle Scholar
  10. Moshiri A, Close J, Reh TA (2004) Retinal stem cells and regeneration. J Dev Biol 48(8–9):1003–1014Google Scholar
  11. Nishiguchi KM, Kaneko H, Nakamura M et al (2008) Identification of photoreceptor precursors in the pars plana during ocular development and after retinal injury. Invest Ophthalmol Vis Sci 49(1):422–428CrossRefPubMedGoogle Scholar
  12. Pignolo RY, Francis MK, Rotemberg MO et al (2003) Putative role for EPC-1/PEDF in the G0 growth arrest of human diploid fibroblasts. J Cell Physiol 195(1):12–20CrossRefPubMedGoogle Scholar
  13. Ramirez-Castillejo C, Sanchez-Sanchez F, Andreu-Agullo C et al (2006) Pigment epithelium-derived factor is a niche signal for neural stem cell renewal. Nat Neurosci 9(3):331–338CrossRefGoogle Scholar
  14. Tombran-Tink J, Chader GG, Johnson LV (1991) PEDF: a pigment epithelium-derived factor with potent neuronal differentiative activity. Exp Eye Res 53:411–414CrossRefPubMedGoogle Scholar
  15. Tropepe V, Coles BL, Chiasson BJ et al (2000) Retinal stem cells in the adult mammalian eye. Science 287:2032–2036CrossRefPubMedGoogle Scholar
  16. Zhao X, Liu J, Ahmad I (2002) Differentiation of embryonic stem cells into retinal neurons. Biochem Biophys Res Commun 297(2):177–84Google Scholar

Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  1. 1.Department of Biomedical SciencesUniversity of Modena and Reggio EmiliaModenaItaly

Personalised recommendations