TY - JOUR T1 - The existence of dead cells in donor corneal endothelium preserved with storage media JF - British Journal of Ophthalmology JO - Br J Ophthalmol SP - 1725 LP - 1730 DO - 10.1136/bjophthalmol-2017-310913 VL - 101 IS - 12 AU - Koji Kitazawa AU - Tsutomu Inatomi AU - Hidetoshi Tanioka AU - Satoshi Kawasaki AU - Hiroko Nakagawa AU - Osamu Hieda AU - Hideki Fukuoka AU - Naoki Okumura AU - Noriko Koizumi AU - Bernie Iliakis AU - Chie Sotozono AU - Shigeru Kinoshita Y1 - 2017/12/01 UR - http://bjo.bmj.com/content/101/12/1725.abstract N2 - Aim To investigate the viability of donor corneal endothelial cells (CECs) preserved in storage media by histological examination.Methods Twenty-eight donor corneas were obtained from SightLife Eye Bank (Seattle, Washington), and redundant peripheral portions of those corneas were used for histological examination after removal of the centre corneal graft for transplantation. To assess cell viability in the corneal endothelium, biostaining experiments were performed using propidium iodide, calcein-AM, Hoechst 33 342, annexin V, anti-vimentin antibody and toluidine blue.Results Histological analysis of the endothelium showed that the cytoplasm of dead cells had low-intensity fluorescence and that their nuclei stained red, while almost all living cells had green cytoplasm and blue-stained nuclei. The mean dead cell rate in the 28 donor corneas was 4.9%±3.3% (mean ±SD) (range: 0.6%–10.5%). The propidium iodide-positive cells stained positive for annexin V, negative for vimentin and pale for toluidine blue. After the specimens were incubated in a culture medium, the red nucleus dead cells dropped off from the level of the blue nucleus living cells.Conclusion Our findings showed the existence of dead cells in storage-media-preserved donor corneal endothelium and that they dropped off after incubation, thus suggesting that the decrease of CECs following keratoplasty may be related to the presence of dead cells. ER -