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Impressum
Preparation of RPE-sheets from porcine bulbi
Gamulescu M. A., Framme C., Roider J.
Augenklinik der Universität Regensburg, Franz-Josef-Strauß-Allee 11, 93053 Regensburg
Objective: The operation of age-related macular degeneration by
extraction of the neovascular membrane is often associated with severe
loss of the retinal pigment epithelium (RPE). In this study, we investigated
a new technique for the preparation of RPE-sheets suitable for transplantation.
Methods: In freshly enucleated porcine eyes (2-4 h after enucleation)
and after removal of the vitreous body and the retina, direct preparation
of RPE is compared to an additional administration of Ca/Mg-free solution
(up to 30 minutes) to losen the adhesion of the RPE at the Bruch membrane.
Following a superficial incision of 3 x 3 mm, the RPE is removed as a
sheet by a subretinal flush of saline.
Results: Examination by light microscopy showed intact RPE-cells
in a coherend single line. With increasing the incubation time with the
Ca/Mgfree solution, we observed an easier preparation of the RPE, however
with the disadvantage of increasing cell loss and discontinuation of the
cell layer. Thus, the optimal incubation time for the preparation of the
RPE was 1-5 min. Despite the fragileness of the prepared RPE-sheets, they
could easily be transferred to another site in the eye globe by means
of a soft instrument. Due to a characteristic reflex, the choroidal and
retinal side were easily identified. We also noted that preparation of
the RPE, when started in the periphery of eye globe, resulted in greater
difficulties in separation and even a damaged RPE layer.
Conclusion: Application of Ca/Mg-free solution may be a suitable
tool for preparation of a functional RPE-sheet that can be dislocated
within the eye globe. However, a time dependence with the application
of Ca/Mg-free solution suggests only a short time window to avoid cellular
and structural damage of the thin cell layer. This technique may be used
for autolog transplantion of a RPE within the eye globe as a therapy for
age-related macular degeneration.