Corneal endothelial or epithelial diseases lead to severe vision impairment. However, transplantation of donor corneae is limited to availability and is impeded by cell loss during tissue processing. Hence, vitrification and cryopreservation of corneal tissue has been under research for a long time (Armitage, 1989; Wusteman et al., 1999), without gaining an alternative to the present usage of organ-cultured or hypothermically stored living tissues (Armitage, 2011). Therefore, the aim in this work was to cryopreserve whole corneas for research purposes and ocular tissue engineering. Methods and Material: Normal porcine corneas were randomly subjected to seven groups: a) Dimethyl sufoxide (DMSO) and Fetal calf serum (FCS), b) 1,2-propanediol (PD) and methyl cellulose (MC), c) 1,2-ethanediol (ED) and hydroxyethyl starch (HES), d) hydroxyethyl cellulose (HEC) and hydroxyethyl starch, e) trehalose (TRE) and povidone (PVP), f) povidone, or g) trehalose. After thawing, corneas were cultured in serum-free medium for three days before subsequent analyses by histology and scanning electron microscope (SEM). Non-frozen corneas were used as controls. Results: Increased stromal thickness was determined for all cryopreserved corneas compared to the controls. Media d and f showed best cryoprotective effects on porcine corneas and supported maintenance of the hexagonal structure of corneal endothelial cells, as visualized by SEM. Conclusion: In this study we established two media suitable to cryopreserve porcine corneal structures.
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