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Jesintha Navaratnam, Eli Gulliksen, Kristine Ustgaard-Andersen, Jon Slettedal, Liv Drolsum, Bjorn Nicolaissen, Aboulghassem Shahdadfar; A Nonsynthetic, Biological Carrier for Cultivated Human Corneal Endothelial Cells (HCECs) for potential therapeutic purposes. Invest. Ophthalmol. Vis. Sci. 2013;54(15):1654.
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The aim of our ongoing study is to establish a carrier for HCECs for therapeutic purposes. In the present study we investigate the feasibility of using nonsynthetic, biological carrier for cultivated HCECs.
Descemet’s membrane with the attached endothelial cells was carefully dissected from human corneas in small strips. One part harvested as non-cultured cells and the other cultivated in corneal endothelial cell growth medium for 6 weeks at 37 degree Celsius with 5% CO2 in a humidified atmosphere and the medium was changed every 2-3 days. The cultivated HCECs were seeded on acellular, nonsynthetic carrier and cultivated for further 3 weeks in corneal endothelial cell expansion medium that was changed every 2-3 days. Cultivated HCECs on nonsynthetic carrier and non-cultured HCEC were comparatively analyzed by qRT-PCR, electron microscopy (EM) and immunohistochemistry.
In our study the cultivated HCECs seeded on nonsynthetic carrier formed a stable monolayer. Our results show that the cultivated HCECs seeded on nonsynthetic carrier and the non- cultured HCECs are functional and express stem cell markers when analyzed by qRT-PCR. The expression levels of markers associated with neural crest, stem cells and corneal endothelial function (SNAI1, SNAI2, SOX9, NES, ZO-1, CX43, VADC2 and VADC3) are higher in cultivated HCECs seeded on nonsynthetic carrier compared to non-cultured HCECs. The structure of cultivated HCECs seeded on nonsynthetic carrier on transmission EM is very similar compared to ex vivo HCECs.
The preliminary results show that nonsynthetic carrier used in this study can potentially be used in therapeutic purposes in the future.
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