Membrane stabilizing effects of calcium and taxol during the cold storage of isolated rat hepatocytes

Background. Calcium plays an important role in liver preservation and prese rvation induces depletion of cellular Ca. This may affect hepatocyte cytosk eleton integrity necessary for maintaining cell shape and organ viability. We tested the effects of a microtubular stabilizer (Taxol) in liver cell pr eservation. Methods. Isolated rat hepatocytes were preincubated with or without a micro tubule stabilizing agent, 100 mu M Taxol at 37 degrees C for 20 min, then s tored in the University of Wisconsin (UW) solution +/-1.5 mM CaCl2 at 4 deg rees C for up to 48 hr. After storage, the cells were rewarmed in Krebs-Hen seleit buffer with air at 37 degrees C for 1 hr. Morphological changes in t he plasma membrane (scanning electron microscopy) and cell viability (perce ntage of lactate dehydrogenase [LDH] release) before and after rewarming we re studied. Results. Nepatocytes showed time-dependent increase in bleb formation (cyto skeleton disruption) during cold storage. Rewarming the cells caused even g reater bleb formation and increased LDH release (cell death). Pretreatment of cells with Taxol and cold storage in the UW solution with 1.5 mM Ca supp ressed both bleb formation and LDH release in 48-hr cold-stored cells. Conclusions. Cold storage of hepatocytes leads to reperfusion injury and ce ll death. This can be suppressed with Taxol and Ca. This suggests that hypo thermia induces changes in cellular Ca and a disruption of the microtubules , leading to loss of cell viability. Improved liver preservation may requir e suppression of Ca-dependent disruption of the cytoskeleton system of live r cells.