June 2013
Volume 54, Issue 15
Free
ARVO Annual Meeting Abstract  |   June 2013
Retinal Ganglion Cell Transplantation After Optic Nerve Injury
Author Affiliations & Notes
  • BO QU
    Bascom Palmer Eye Institute and Interdisciplinary Stem Cell Institute, University of Miami, miami, FL
    The 4th Affiliated Hospital of China Medical University, Shenyang, China
  • Jonathan Hertz
    Bascom Palmer Eye Institute and Interdisciplinary Stem Cell Institute, University of Miami, miami, FL
  • Roshni Patel
    Bascom Palmer Eye Institute and Interdisciplinary Stem Cell Institute, University of Miami, miami, FL
  • Yan Wang
    Bascom Palmer Eye Institute and Interdisciplinary Stem Cell Institute, University of Miami, miami, FL
  • Jeffrey Goldberg
    Bascom Palmer Eye Institute and Interdisciplinary Stem Cell Institute, University of Miami, miami, FL
  • Footnotes
    Commercial Relationships BO QU, None; Jonathan Hertz, None; Roshni Patel, None; Yan Wang, None; Jeffrey Goldberg, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science June 2013, Vol.54, 2243. doi:
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    • Get Citation

      BO QU, Jonathan Hertz, Roshni Patel, Yan Wang, Jeffrey Goldberg; Retinal Ganglion Cell Transplantation After Optic Nerve Injury. Invest. Ophthalmol. Vis. Sci. 2013;54(15):2243.

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

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Abstract
 
Purpose
 

To compare the long term transplantation of acutely purified retinal ganglion cells (RGCs) onto normal and optic nerve crushed retina both ex vivo and in vivo.

 
Methods
 

Optic nerve crush was performed on Sprague-Dawley rats. RGCs from postnatal 4 (P4) transgenic β-actin-GFP mice were acutely purified by sequential immunopanning, transplanted onto retina explants at different time points and co-cultured for 7days. Also, RGCs were injected intravitreally and animals survived for 28 days. After immunostaining, transplanted RGC survival was quantified by nuclear morphology. Migration into the ganglion cell layer was quantified with 3D Z-stack projections using confocal fluorescence microscopy.

 
Results
 

Ex vivo transplanted RGCs to the 21, 30, and 41 days-post-axotomy damaged retina survived, migrated, and extended neurites more than controls or 14 days-post axotomy, at increasing times after optic nerve crush. The transplanted RGCs gave much more neurites and migrated deep into the retina, and also we found more cells clustered. In vivo, more cells survived in the 30 day-post axotomy eyes. About 35% of transplanted RGCs migrated under the ganglion cell layer or even into inner nuclear, compared to about 6% in normal eyes. In the inner retinal layers, donor RGCs demonstrated typical RGC morphology, extending long axons and short dendrites, in the previously axotomized but rarely in normal eyes.

 
Conclusions
 

After optic nerve injury, transplanted purified RGCs can survive and migrate better both ex vivo and in vivo compared to transplant into normal eyes. Ex vivo, donor cells elaborate more neurites in shorter times, whereas in vivo, cells migrate deeper and demonstrate more typical neurite morphology.

 
 
retina explant immunostaining of 30 day-post axotomy eyes 7days after RGC transplantation ex vivo. GFP in green, βIII-tubulin in red, Dapi in blue. Many GFP positive cells migrate into retina, extending lots of neurites and many cells cluster.
 
retina explant immunostaining of 30 day-post axotomy eyes 7days after RGC transplantation ex vivo. GFP in green, βIII-tubulin in red, Dapi in blue. Many GFP positive cells migrate into retina, extending lots of neurites and many cells cluster.
 
 
retina explant immunostaining of 30 day-post axotomy eyes 28days after RGC transplantation in vivo. GFP in green, βIII-tubulin in red, Dapi in blue. Many GFP positive cells migrate into retina, some of them extend long neuritis.
 
retina explant immunostaining of 30 day-post axotomy eyes 28days after RGC transplantation in vivo. GFP in green, βIII-tubulin in red, Dapi in blue. Many GFP positive cells migrate into retina, some of them extend long neuritis.
 
Keywords: 531 ganglion cells • 741 transplantation • 494 degenerations/dystrophies  
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