Purchase this article with an account.
M. Halberstadt, B. Frueh, M. Kilchenmann, F. Flückiger, M. Hagenah; Cryopreservation of Organ Cultured Human Donor Corneas . Invest. Ophthalmol. Vis. Sci. 2005;46(13):4529.
Download citation file:
© ARVO (1962-2015); The Authors (2016-present)
Purpose: To assess the impact of organ culture time prior to cryopreservation on freeze–thaw induced endothelial cell loss. Thereby, evaluating the possibility of cryopreservation of human corneas as a method to extend storage time after organ culturing. Methods: Twenty–seven human donor corneas who had been organ cultured for up to 42 days (mean 22.8 ± 12.9) were cryopreserved in minimal essential medium containing 10% dextran with a molecular weight of 500,000 as an extracellular cryoprotectant, at a cooling rate of 1°C/min and then stored in liquid nitrogen at –196°C. After thawing, the tissue was again organ cultured to detect latent cell damage. Results: After cryopreservation and subsequent organ culturing, the endothelial cell density dropped by 30.5% to 1404 ± 483 cells/mm2, and 2 of the corneas had a completely necrotic endothelium (p= 0.002). Univariate analysis revealed an organ culture period of more than 28 days to have a significant impact on endothelial cell loss after thawing (p< 0.001). Variables like duration of deswelling period in dextran (p= 0.929), post–mortem time (p= 0.727), cell loss during organ culture prior to freezing (p= 0.178), and age (p= 0.689) revealed to have no significant impact on freeze–thaw induced endothelial cell loss. Conclusions: After an organ culture period of up to 3 weeks the tissue can still be cryopreserved and stored for an indefinite time without a significant increase of endothelial cell loss.
This PDF is available to Subscribers Only