Evaluation of Megacell MEM as a storage medium for corneas destined for transplantation.
OBJECTIVE
Gibco's Minimum Essential Medium with Earle's salts and HEPES supplemented with glutamine, antibiotics (EB MEM) and 2% foetal calf serum (FCS) is used in European eye banks to store corneas. Although FCS is important to endothelial cell survival in this medium, it is a potential biohazard. Megacell MEM, formulated to reduce the FCS requirement of cells by a factor of 5, has therefore been evaluated as a corneal storage medium.


METHODS
Corneal stromal and epithelial cells were incubated in Megacell MEM (serum-free or 2% FCS) to assess their viability in these media. Endothelial cell densities of paired corneas held in either EB MEM 2% FCS or Megacell MEM (serum-free or 2% FCS) were measured over 5 weeks. Discs subsequently punched from the centre of these corneas were weighed, dried and reweighed to determine hydration levels.


RESULTS
Both corneal stromal and epithelial cells proliferated in Megacell MEM 2% FCS. Relative to EB MEM, 2% FCS Megacell MEM prolonged the viability of corneal endothelial cells and improved their morphological appearance, irrespective of whether it contained FCS or not. This was independent of corneal swelling.


CONCLUSIONS
Serum-free Megacell MEM is a better storage medium than EB MEM 2% FCS for corneas destined for transplantation.
Smith
Valerie A
VA
University of Bristol, Academic Unit of Ophthalmology, Bristol Eye Hospital, Bristol, UK. Val.Smith@bristol.ac.uk
Johnson
Terrell
T
D000293:Adolescent; D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D000818:Animals; D002452:Cell Count; D049109:Cell Proliferation / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D003319:Corneal Stroma / Q000166:cytology / Q000187:drug effects* / Q000254:growth & development; D016039:Corneal Transplantation / Q000379:methods*; D007562:Creutzfeldt-Jakob Syndrome / Q000209:etiology; D016895:Culture Media, Serum-Free / Q000737:chemistry / Q000494:pharmacology*; D019573:Epithelium, Corneal / Q000166:cytology / Q000187:drug effects* / Q000254:growth & development; D005125:Eye Banks / Q000379:methods; D006801:Humans; D008875:Middle Aged; D009924:Organ Culture Techniques / Q000379:methods; D009926:Organ Preservation / Q000379:methods; D019188:Organ Preservation Solutions / Q000009:adverse effects / Q000737:chemistry / Q000494:pharmacology*
D016895:Culture Media, Serum-Free; D019188:Organ Preservation Solutions