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EPITHELIAL DEFECTS IN CORNEAS IN ORGAN CULTURE
CTS-Manchester Eye Bank
& Ophthalmic Pathology Service
Isaac Zambrano
Fiona Carley
Luciane Irion
Richard Bonshek
Taking the eye out of the pot
Cleaning the remains of the tissue
Washing eyes in saline, iodine & Sodium thiosulphate
Cutting sclera with trephine
Excision of corneoscleral disc using scalpel
Inserting suture in sclera
Cleaning of eyes and excision of corneas
Cornea - organ culture (31 Cornea - organ culture (31 to 37°C)to 37°C)
Culture medium: Supply electrolytes, nutrients, Culture medium: Supply electrolytes, nutrients, vitamins, hormones, proteins vitamins, hormones, proteins Maintain metabolismMaintain metabolism Support cell proliferation and protein synthesisSupport cell proliferation and protein synthesis Support cellular repair mechanismsSupport cellular repair mechanisms
Placing corneas in organ culturePlacing corneas in organ culture
Placing the corneoscleral disc in organ culture
Incubating for up to 28
days at 34 0C
Issuing corneasIssuing corneas
Release from quarantineRelease from quarantine 3 days before scheduled date of graft, corneal 3 days before scheduled date of graft, corneal
endothelium examined by light microscopyendothelium examined by light microscopy Estimate cell density (minimum 2200 cells/mm²)Estimate cell density (minimum 2200 cells/mm²) Dead/missing cells (trypan blue staining)Dead/missing cells (trypan blue staining) OpacitiesOpacities
If suitable for PK, cornea placed into medium If suitable for PK, cornea placed into medium containing 5% dextran T500 to reverse containing 5% dextran T500 to reverse stromal oedema and returned to 34°Cstromal oedema and returned to 34°C
Cornea despatched at ambient temperature to Cornea despatched at ambient temperature to recipient hospital day before graftrecipient hospital day before graft
3 Cases
Corneas stored in culture
Epithelial defects before or after culture with dextran medium
FCY-13A.
75 years old male.
Death to enucleation: 15hrs
Enucleation to culture: 17hrs
Death to culture: 33hrs
Cause of death: Pneumonia, Myocardial infarction
FCY13-A
FCY-13A
FCY-3A
60 years old male.
Death to enucleation: 20hrs
Enucleation to culture: 8hrs
Death to culture: 28hrs
Cause of death: alcoholic liver disease
FCY-3A
FCY-3A
FCY-3A
FCY-3A
FCY-1B
60 years old female
Death to donation: 19hrs
Enucleation to culture: 18hrs
Death to culture: 38hrs
Cause of death: Ca Lung
FCY1-A
FCY1-B
FCY1
FCY1
SUMMARY
Several cases of epithelial defects in corneas stored in organ culture
Often visible only when corneas are transferred to culture medium with dextran
Epithelial defects are of non-infectious nature
No correlation with time of death to enucleation and to storage, age, endothelial cell count or cause of death
Why some corneas develop such abnormalities?