Poloxamer 188 enhances endothelial cell survival in bovine corneas in cold storage.

Purpose: 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.

Long-term potentiation of exocytosis and cell membrane repair in fibroblasts.

We previously found that a microdisruption of the plasma membrane evokes Ca(2+)-regulated exocytosis near the wound site, which is essential for membrane resealing. We demonstrate herein that repeated membrane disruption reveals long-term potentiation of Ca(2+)-regulated exocytosis in 3T3 fibroblasts, which is closely correlated with faster membrane resealing rates. This potentiation of exocytosis is cAMP-dependent protein kinase A dependent in the early stages (minutes), in the intermediate term (hours) requires protein synthesis, and for long term (24 h) depends on the activation of cAMP response element-binding protein (CREB).