ArticleThe British journal of ophthalmology2002
Value of two mortality assessment techniques for organ cultured corneal endothelium: trypan blue versus TUNEL technique.
Article in The British journal of ophthalmology, 2002. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 22 papers, 1 of them a synthesis that pooled it.
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22 citing papers in PubMed, 1 synthesis or guideline pooled it, 87 citations in OpenAlex.
- A systematic review of ultrasound-mediated drug delivery to the eye and critical insights to facilitate a timely path to the clinic.Theranostics · 2023Pooled it
- Trial
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- The impact of umbilical cord blood platelet lysate on human corneal endothelium during organ culture.International ophthalmology · 2026Article
- Optimized alveolar epithelial cell model for chroniciScience · 2025Article
- A corneal and whole eye globe bovine ex vivo model to mimic human donor corneal storage conditions and eye surgeries.Cell and tissue banking · 2025Article
- Expression of Yes-associated protein in endothelial cells of human corneas before and after storage in organ culture.Scientific reports · 2024Article
- Retrospective Clinical Outcomes of Keratoplasty Using Human Donor Corneas Preserved in Eusol-C Hypothermic Storage Medium.Journal of clinical medicine · 2024Article
- Effects of Intracameral Drugs and Dyes on Corneal Endothelial Cell Apoptosis in a Rat Model: AnTurkish journal of ophthalmology · 2022Article
- Specular Microscopy of Human Corneas Stored in an Active Storage Machine.Journal of clinical medicine · 2022Article
- Surface quality and endothelial cell viability after femtosecond laser-assisted donor lenticule preparation for endothelial keratoplasty - AnIndian journal of ophthalmology · 2020Article
- Apoptosis Induction and Alteration of Cell Adherence in Human Lung Cancer Cells under Simulated Microgravity.International journal of molecular sciences · 2019Article
- [Critical endothelial procedures during posterior lamellar graft preparation and transplantation].Der Ophthalmologe : Zeitschrift der Deutschen Ophthalmologischen Gesellschaft · 2017Review
- Real-time assessment of corneal endothelial cell damage following graft preparation and donor insertion for DMEK.PloS one · 2017Article
- Global cell-by-cell evaluation of endothelial viability after two methods of graft preparation in Descemet membrane endothelial keratoplasty.The British journal of ophthalmology · 2016Article
- Preservation of Preloaded DMEK Lenticules in Dextran and Non-Dextran-Based Organ Culture Medium.Journal of ophthalmology · 2016Article
- Impact of temporary hyperthermia on corneal endothelial cell survival during organ culture preservation.Graefe's archive for clinical and experimental ophthalmology = Albrecht von Graefes Archiv fur klinische und experimentelle Ophthalmologie · 2015Article
- Anti-apoptotic gene therapy prolongs survival of corneal endothelial cells during storage.Gene therapy · 2011Article
- Corneal graft rejection occurs despite Fas ligand expression and apoptosis of infiltrating cells.The British journal of ophthalmology · 2005Article
- Mechanisms of staurosporine induced apoptosis in a human corneal endothelial cell line.The British journal of ophthalmology · 2003Article
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9 authors at 1 institution in 2 countries.
Funding
No grant is acknowledged in the PubMed record.
Abstract
BACKGROUND/
aimIt is known that trypan blue staining is not a good predictor of loss of corneal endothelial cells (ECs) during organ culture. As it is primarily an indicator of membrane integrity, it would also not be expected to identify ECs undergoing apoptosis. The aim of this study was to determine the ability of the in situ TdT dUTP mediated nick end labelling (TUNEL) technique to detect cell death in the corneal endothelium caused by apoptosis during organ culture, compared with conventional vital staining with trypan blue.
methods31 human corneas were organ cultured at 31C for 3-35 days. Staurosporine was used to induce apoptosis in five control corneas. The endothelium was assessed by trypan blue and by the in situ TUNEL technique. The percentages of trypan and TUNEL positive ECs were compared. Their links with sex, donor age, time from donor death and organ culture, initial and final EC density and cell loss were studied.
resultsTUNEL stained ECs were observed in all corneas. TUNEL positive ECs were mostly located either in corneal folds or at the periphery of corneal folds showing central shedding. The mean percentage of cell death at the end of storage, assessed by the trypan blue technique, was 1.47% (SD 2.63, range 0.03-12); assessed by the TUNEL technique it was 12.7% (SD 16.4 range 0.6-65.5). There was a significant correlation between the two techniques (r = 0.7, p<0.001). The percentage of TUNEL stained ECs was correlated negatively with EC density at the end of storage (r = -0.47, p <0.005) and positively with percentage EC loss during storage (r = 0.46, p < 0.05).
conclusionThis study demonstrates that organ cultured corneas systematically carry non-viable ECs that are implicated in cell death by apoptosis and go undetected when trypan blue staining is used. Because the in situ TUNEL assay detects earlier events in the cell death process than does trypan blue, it should be used to quantify endothelial viability, especially for experiments with new storage media.
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