May 2004
Volume 45, Issue 13
Free
ARVO Annual Meeting Abstract  |   May 2004
Organization and differentiation of in vitro expanded retinal progenitor cells post–transplantation into degenerate rats
Author Affiliations & Notes
  • C.H. Mui
    Doheny Eye Institute/Ophthalmolo, Doheny Retina Institute, Keck School of Medicine USC, Los Angeles, CA
  • G.T. Qiu
    Doheny Eye Institute/Ophthalmolo, Doheny Retina Institute, Keck School of Medicine USC, Los Angeles, CA
  • M.J. Seiler
    Doheny Eye Institute/Ophthalmolo, Doheny Retina Institute, Keck School of Medicine USC, Los Angeles, CA
    Cell & Neurobiology, Keck School of Medicine, USC, Los Angeles, CA
  • S. Arai
    Doheny Eye Institute/Ophthalmolo, Doheny Retina Institute, Keck School of Medicine USC, Los Angeles, CA
    Division of Ophthalmology and Vision Science, Graduated School of Medical and Dental Sciences, Niigatta, Japan
  • R.B. Aramant
    Anatomical Science & Neurobiology, Univ. Louisville School of Medicine, Louisville, KY
  • E. de Juan
    Doheny Eye Institute/Ophthalmolo, Doheny Retina Institute, Keck School of Medicine USC, Los Angeles, CA
  • S.R. Sadda
    Doheny Eye Institute/Ophthalmolo, Doheny Retina Institute, Keck School of Medicine USC, Los Angeles, CA
  • Footnotes
    Commercial Relationships  C.H. Mui, None; G.T. Qiu, None; M.J. Seiler, None; S. Arai, None; R.B. Aramant, None; E. de Juan, None; S.R. Sadda, None.
  • Footnotes
    Support  NIH EY 03040
Investigative Ophthalmology & Visual Science May 2004, Vol.45, 5403. doi:
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      C.H. Mui, G.T. Qiu, M.J. Seiler, S. Arai, R.B. Aramant, E. de Juan, S.R. Sadda; Organization and differentiation of in vitro expanded retinal progenitor cells post–transplantation into degenerate rats . Invest. Ophthalmol. Vis. Sci. 2004;45(13):5403.

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      © ARVO (1962-2015); The Authors (2016-present)

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Abstract

Abstract: : Purpose:To characterize the organization and differentiation of retinal progenitor cells (RPCs) following transplantation into the vitreous or subretinal space of S334ter5 rhodopsin mutant transgenic (slow retinal degeneration) rats. Materials and Methods:RPCs were isolated from human placental alkaline phosphatase (hPAP)–positive embryonic day 17 (E17) rat retina and expanded in serum–free defined media supplemented with epidermal growth factor (EGF) and basic fibroblast growth factor (bFGF). RPCs at passage 2 (P2) were identified by double immunostaining with nestin and neuronal or glial cell markers. Cultured P2 retinal progenitor cells were transplanted into postnatal day 17 (P17) S334ter5 rats via intravitreal or subretinal injection. Four weeks after transplantation, the eyes were studied microscopically with immunohistochemistry for various retinal cell specific markers. Results:Before transplantation, the in vitro RPCs co–expressed nestin with MAP–2. Four weeks after transplantation, the P2 RPC–derived (hPAP+) cells were found to organize into sheets and integrate into the diseased host retina. Grafted cells were found to express multiple retinal specific neuronal markers including: rhodopsin, protein kinase C, calbindin, and recoverin. Regardless of the location of the transplant (subretinal versus intravitreal), approximately 50% of the grafted cells expressed photoreceptor–specific markers, whereas less than 2% expressed Muller/glial specific markers such as GFAP and CRALBP. Conclusions:In contrast to previous studies depicting the formation of cell clumps or neural spheres, RPCs transplanted into retinal degenerate rats by our technique yielded well–organized sheets of cells. In addition, a large proportion of these cells developed phenotypic characteristics of photoreceptors, regardless of the transplantation method (intravitreal versus subretinal), suggesting that intrinsic signals may contribute significantly to RPC differentiation over the extrinsic (environmental) cues. Supported by: Foundation Fighting Blindness, Anonymous Sponsor, Foundation for Retinal Research, Fletcher Jones Foundation, NIH EY03040.

Keywords: transplantation • retina • retinal culture 
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