Poloxamer 188 enhances endothelial cell survival in bovine corneas in cold storage.
OBJECTIVE
To improve corneal endothelial cell survival during cold preservation by the addition of a compound that enhances cell membrane repair.


METHODS
Cell survival was assessed using intracellular accumulation of the fluorescent molecule calcein as a marker for endothelial cell viability in bovine corneas stored at 4 degrees C in Optisol-GS. The effect on endothelial cell survival of artificially lowering the surface tension of the cell membrane was also assayed by the addition of a surfactant, Poloxamer 188 (PLURONIC F68), which has been shown to reseal damaged cell membranes, even under conditions where metabolism is inhibited.


RESULTS
Endothelial cell survival in corneas stored at 4 degrees C in Optisol-GS was increased by the addition of 1 mg/mL Poloxamer 188. The level of dead or missing corneal endothelial cells was less than 1% for corneas stored for 6, 9, or 12 days in Optisol-GS with 1 mg/mL Poloxamer 188 at 4 degrees C. In contrast, dead or missing corneal endothelial cell percentages were increased to approximately 4% after 9 days and approximately 10% after 12 days in storage in Optisol-GS without Poloxamer 188 at 4 degrees C.


CONCLUSIONS
The surfactant Poloxamer 188 enhances endothelial cell survival in cold-stored bovine corneas.
Steinhardt
Richard Anthony
RA
Department of Molecular and Cell Biology, University of California, Berkeley, CA 94720-3200, USA. ricksteinhardt@berkeley.edu
Alderton
Janet Marilyn
JM
D000818:Animals; D002417:Cattle; D002452:Cell Count; D002470:Cell Survival / Q000187:drug effects; D002809:Chondroitin Sulfates / Q000494:pharmacology; D003080:Cold Temperature; D045424:Complex Mixtures / Q000494:pharmacology; D016895:Culture Media, Serum-Free / Q000494:pharmacology; D003911:Dextrans / Q000494:pharmacology; D004728:Endothelium, Corneal / Q000166:cytology* / Q000187:drug effects*; D005452:Fluoresceins; D005456:Fluorescent Dyes / Q000494:pharmacology; D005839:Gentamicins / Q000494:pharmacology; D008856:Microscopy, Fluorescence; D019188:Organ Preservation Solutions; D020442:Poloxamer / Q000494:pharmacology*; D013501:Surface-Active Agents / Q000494:pharmacology*
D045424:Complex Mixtures; D016895:Culture Media, Serum-Free; D003911:Dextrans; D005452:Fluoresceins; D005456:Fluorescent Dyes; D005839:Gentamicins; D019188:Organ Preservation Solutions; D013501:Surface-Active Agents; D020442:Poloxamer; D002809:Chondroitin Sulfates; C075736:Optisol; C007740:fluorexon