RT Journal Article
SR Electronic
T1 The existence of dead cells in donor corneal endothelium preserved with storage media
JF British Journal of Ophthalmology
JO Br J Ophthalmol
FD BMJ Publishing Group Ltd.
SP 1725
OP 1730
DO 10.1136/bjophthalmol-2017-310913
VO 101
IS 12
A1 Koji Kitazawa
A1 Tsutomu Inatomi
A1 Hidetoshi Tanioka
A1 Satoshi Kawasaki
A1 Hiroko Nakagawa
A1 Osamu Hieda
A1 Hideki Fukuoka
A1 Naoki Okumura
A1 Noriko Koizumi
A1 Bernie Iliakis
A1 Chie Sotozono
A1 Shigeru Kinoshita
YR 2017
UL http://bjo.bmj.com/content/101/12/1725.abstract
AB 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.