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Constandinos Carserides, Rachel Williams, Vito Romano, Hannah Jane Levis; Development of an ex vivo organ culture model of the cornea: a useful tool for corneal studies.. Invest. Ophthalmol. Vis. Sci. 2019;60(9):2185.
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© ARVO (1962-2015); The Authors (2016-present)
We have developed an ex vivo organ culture model of the cornea that can be used for a range of corneal related studies.
Paired freshly enucleated porcine corneas were mounted on an artificial anterior chambers (AACs) and cultured under short term (ST, 24 hour culture) and long term (LT, 5 day culture) conditions. For ST culture AACs were contained within an acrylic box and placed in an incubator at 31oC. For LT culture AACs were contained within an aerated acrylic box and placed in an incubator at 27oC. Under both conditions the epithelial surface of the corneas were automatically moistened with a droplet of epithelial cell culture media at a flow rate of 50-200µl/hr with the use of a syringe pump. Additionally, the endothelial surface was manually perfused with fresh endothelial cell culture media. One cornea from each pair was then damaged by scraping of the endothelium. Central corneal thickness (CCT) was measured using pachymetry or optical coherence tomography (OCT).
Mean thickness of paired whole porcine globes using pachymetry was measured at 918±64 µm (n=6). Under ST culture conditions, damaged corneas increased in mean thickness by 62±3% (n=3) when compared to the undamaged cornea from the same donor which was measured by OCT. Under LT culture, paired corneas were successfully cultured for 5 days with no evidence of contamination or tissue decomposition. Pachymetery indicated that damaged corneas significantly increased in CCT, this occurred as rapidly as 1 day in culture. In comparison, daily pachymetry measurements were obtained for undamaged corneas with a delayed increase in thickness (Day1= 930±19µm to Day 5= 1410±16µm, n=1) which occurred over a 5 day period and clear corneal opacity was observed.
Automated moistening of the epithelium maintained a detectable CCT up to 5 days in culture. Moderate swelling occured, however, pachymetry indicates that this is still significantly lower when compared to damaged corneas. Continuous work will involve introducing a flow of endothelial culture media into the anterior chamber to maintain a stable pressure which can be detected with rebound tonometry. Tissue morphology will be evaluated by histological analysis and cell morphology assesed by alizarin red staining.
This abstract was presented at the 2019 ARVO Annual Meeting, held in Vancouver, Canada, April 28 - May 2, 2019.
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