Ex vivo evaluation of intravitreal mesenchymal stem cell viability using bioluminescence imaging.

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