A sensitive method for testing the quality of organ culture media and of individual medium components in a cornea bank.
BACKGROUND
It has been suggested that variations in the quality of organ culture preservation media are responsible for variations in early postoperative graft morphology. Spates of such variations have been observed repeatedly for short periods. This paper reports the results of a series of grafts with low postoperative clearing observed during a period of 6 weeks. Simultaneously, preoperative phase-contrast microscopy evaluation of the corneal endothelium revealed that an unusually large proportion of donor corneae were unsuitable for transplantation.


METHODS
The corneal storage media were therefore rigorously screened, paying particular attention to specific components and properties of the medium, including L-glutamine, amphotericin B, water quality, pH, and the glassware used. Possible toxic effects were identified by means of a sensitive growth assay performed using isolated human corneal endothelial cells.


RESULTS
The evaluation demonstrated that both the water quality and the L-glutamine which had been used for preparation of the medium were substandard during the period in which poor clinical results were obtained.


CONCLUSIONS
It is recommended that cornea banks undertaking long-term organ culture use standardized protocols and carefully monitored equipment. The quality of the basal media and supplements should be routinely checked.
Engelmann
K
K
Universitäts-Augenklinik, Hamburg, Germany.
Sobottka Ventura
A
A
Drexler
D
D
Staude
H J
HJ
D000666:Amphotericin B / Q000032:analysis*; D000818:Animals; D000900:Anti-Bacterial Agents / Q000032:analysis*; D016039:Corneal Transplantation / Q000473:pathology; D017077:Culture Media, Conditioned / Q000737:chemistry / Q000592:standards*; D004728:Endothelium, Corneal / Q000187:drug effects / Q000254:growth & development; D005125:Eye Banks; D005973:Glutamine / Q000032:analysis*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D009924:Organ Culture Techniques; D009926:Organ Preservation; D012680:Sensitivity and Specificity; D013552:Swine; D014019:Tissue Donors; D014867:Water / Q000032:analysis*
D000900:Anti-Bacterial Agents; D017077:Culture Media, Conditioned; D014867:Water; D005973:Glutamine; D000666:Amphotericin B