April 2014
Volume 55, Issue 13
Free
ARVO Annual Meeting Abstract  |   April 2014
Ex vivo evaluation of intravitreal mesenchymal stem cell viability using bioluminescence imaging.
Author Affiliations & Notes
  • Marcelo Jordão L Silva
    Opthalmology, FMRP University of São Paulo, Ribeirão Preto, Brazil
  • Priscila C Ferreira
    Opthalmology, FMRP University of São Paulo, Ribeirão Preto, Brazil
  • Rubens C Siqueira
    Opthalmology, FMRP University of São Paulo, Ribeirão Preto, Brazil
  • Rodrigo Jorge
    Opthalmology, FMRP University of São Paulo, Ribeirão Preto, Brazil
  • Andre Messias
    Opthalmology, FMRP University of São Paulo, Ribeirão Preto, Brazil
  • Maria Lourdes Veronese Rodrigues
    Opthalmology, FMRP University of São Paulo, Ribeirão Preto, Brazil
  • Rodrigo J Calado
    Opthalmology, FMRP University of São Paulo, Ribeirão Preto, Brazil
  • Dimas T Covas
    Opthalmology, FMRP University of São Paulo, Ribeirão Preto, Brazil
  • Jayter Silva Paula
    Opthalmology, FMRP University of São Paulo, Ribeirão Preto, Brazil
  • Footnotes
    Commercial Relationships Marcelo Silva, None; Priscila Ferreira, None; Rubens Siqueira, None; Rodrigo Jorge, None; Andre Messias, None; Maria Rodrigues, None; Rodrigo Calado, None; Dimas Covas, None; Jayter Paula, None
  • Footnotes
    Support None
Investigative Ophthalmology & Visual Science April 2014, Vol.55, 1370. doi:
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      Marcelo Jordão L Silva, Priscila C Ferreira, Rubens C Siqueira, Rodrigo Jorge, Andre Messias, Maria Lourdes Veronese Rodrigues, Rodrigo J Calado, Dimas T Covas, Jayter Silva Paula; Ex vivo evaluation of intravitreal mesenchymal stem cell viability using bioluminescence imaging.. Invest. Ophthalmol. Vis. Sci. 2014;55(13):1370.

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

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Abstract
 
Purpose
 

Bone marrow-derived mesenchymal stem cells (MSC) therapy is a promising treatment for several degenerative diseases, including retinopathies and glaucoma, however no reproducible method of monitoring these cells into the eye has been established. The aim of this study was to describe a successful bioluminescence imaging (BLI) to detect viable luciferase-expressing MSC into the eye.

 
Methods
 

Human donors’ MSC in culture were infected with 50ul of luciferase lentiviral vector (3,000 viral particles/cell) prior to intraocular injections. Three eyes of two rabbits were evaluated through BLI after receiving 1x106 luciferase-expressing MSC intravitreally with (E1) or without (E2) D-luciferin (40mg/ml - 300 µL of PBS), via pars plana. D-luciferin (40mg/ml - 300 µL of PBS) without cells was injected in a third eye at beginning (E3) and after one hour in E2.

 
Results
 

E1 showed high BLI radiance report and decay in eight hours. After D-luciferin infusion, E2 also displayed high average radiance, with similar decay rate of E1. No signal was observed in E3. Figure 1 show the bioluminescence imaging acquired from the three experimental eyes and the figure 2 show the distribution of total amount of captured photons from the three eyes, using bioluminescence, during the eight hours period.

 
Conclusions
 

Identification of cell location and viability is still an important problem regarding the use of MSC for eye diseases. This is maybe the first ex vivo study demonstrating BLI is useful and reliable method to address these issues.

     
Keywords: 721 stem cells • 561 injection • 449 cell survival  
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