epithelial defects in corneas in organ culture

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?