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Br J Ophthalmol. 2017 Dec;101(12):1725-1730. doi: 10.1136/bjophthalmol-2017-310913. Epub 2017 Oct 5.

The existence of dead cells in donor corneal endothelium preserved with storage media.

Author information

1
Department of Frontier Medical Science and Technology for Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
2
Baptist Eye Institute, Kyoto, Japan.
3
Department of Ophthalmology, Kyoto Prefectural University of Medicine, Kyoto, Japan.
4
Department of Ophthalmology, Osaka University, Suita, Japan.
5
Department of Biomedical Engineering, Faculty of Life and Medical Sciences, Doshisha University, Kyotanabe, Japan.
6
SightLife, Seattle, Washington, DC, USA.

Abstract

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.

KEYWORDS:

corneal endothelial cell; corneal endothelium; corneal transplantation; donor cornea; keratoplasty; storage media

[Indexed for MEDLINE]

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