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The 8‐fold quadrant dissection method for ex vivo human interventional retinal experimentation
Author(s) -
Murali A.,
RamloganSteel C.,
Andrzejewski S.,
Dhungel B.,
Steel J.,
Layton C.
Publication year - 2017
Publication title -
acta ophthalmologica
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.534
H-Index - 87
eISSN - 1755-3768
pISSN - 1755-375X
DOI - 10.1111/j.1755-3768.2017.01558
Subject(s) - retina , quadrant (abdomen) , retinal , ex vivo , ophthalmology , dissection (medical) , population , medicine , biology , in vivo , anatomy , pathology , neuroscience , microbiology and biotechnology , environmental health
Purpose Retinal research relies on animal and in‐vitro models which lack many of the characteristics of human retina. We have instead established a reproducible ex‐vivo model of primary retinal explants derived from human donor eye cups. Methods A dissection strategy performed independently by two investigators was designed to maximize retinal tissue whilst maintaining experimentally reproducible fragments of retina suitable for experimental purposes. The retina was divided into 4 quadrants through the fovea and equivalent distribution of photoreceptors between quadrants was confirmed with CD 73 staining in 7 pairs of donor eyes with flow cytometry. In the 8 quadrants from each of the 7 donors, the standard error in proportion of photoreceptors was 0.8–2.3%. Results Cellular composition of free floating retinal explants were followed for 2 months in 10 retinas with quadrant dissection and 8 with random dissection. Explants could be maintained for 2 months with live populations of photoreceptors, ganglion cells and Müller cells detected by flow cytometry. In contrast, amacrine and horizontal cells, decreased by 90% at 7 days. Only quadrant dissection from the same patient showed reproducible and reliable proportions of cell populations between dissections at any time point. There was no statistically significant relationship between the proportion of any cell population and donor age, time after death, time to storage or time in storage. Conclusions The 8 fold quadrant dissection method from a single donor forms an attractive human experimental model for interventional testing in retinal research. We demonstrate the utility of this model in a simple neuroprotection study, showing that insulin protected against CoCl 2 induced hypoxia in human photoreceptors.