Abstract
Purpose :
We have previously reported that the intravenous injection of bone marrow-derived cells (BMDC) programmed with the RPE65 gene promoted visual recovery in a mouse model of AMD. Because of that, the aim of this study was to characterize the spatial and temporal recruitment of those programmed cells to the RPE layer.
Methods :
2-month old C57BL/6J female mice received a subretinal injection of AAV1-SOD2 ribozyme to induce the AMD mouse model. One month after the injection, systemic intravenous injection of 50,000 GFP+ RPE65-programmed mouse bone marrow-derived cells (BMDC)/animal was performed. Animals were terminated at different time-points from one to 60 minutes after cell injection and the localization of GFP+ cells was determined by fluorescence microscopy in RPE/retinal flat mounts and sections.
Results :
The analysis of RPE/retina flat mounts showed that the majority of the GFP+ cells were found exclusively in the RPE layer. In the first two time-points, the cells were found especially in the proximity of ora serrata (peripheral) region of the RPE flat mount while few cells were observed in the center, near the optic nerve. In the following time-points, up to one hour after cell injection, the number of cells in the peripheral area was reduced while in the central area the number was increased. The neural retina showed cells scattered in the center of the tissue that presented a higher density in the first 2 minutes which decreased in the following time-points after cell injection. These findings were confirmed by immunofluorescence analysis of retinal sections, where we observed individual cells expressing both GFP and RPE65 markers.
Conclusions :
In this study our group shows that the reprogrammed BMDC, administrated systemically, are capable of migrating to the eye, enter the retina via both the central retina and posterior ciliary arteries and migrate to the injured RPE layer.
This is a 2021 ARVO Annual Meeting abstract.