Membrane stabilizing effects of calcium and taxol during the cold storage of isolated rat hepatocytes
Kim, J.S.; Southard, J.H.
Transplantation 68(7): 938-943
1999
ISSN/ISBN: 0041-1337 PMID: 10532531 Document Number: 499717
Background: Calcium plays an important role in liver preservation and preservation induces depletion of cellular Ca. This may affect hepatocyte cytoskeleton integrity necessary for maintaining cell shape and organ viability. We tested the effects of a microtubular stabilizer (Taxol) in liver cell preservation. Methods: Isolated rat hepatocytes were preincubated with or without a microtubule stabilizing agent, 100 muM Taxol, at 37degreeC for 20 min, then stored in the University of Wisconsin (UW) solution +- 1.5 mM CaCl2 at 4degreeC for up to 48 hr. After storage, the cells were rewarmed in Krebs-Henseleit buffer with air at 37degreeC for 1 hr. Morphological changes in the plasma membrane (scanning electron microscopy) and cell viability (percentage of lactate dehydrogenase (LDH) release) before and after rewarming were studied. Results: Hepatocytes showed time-dependent increase in bleb formation (cytoskeleton disruption) during cold storage. Rewarming the cells caused even greater bleb formati on and increased LDH release (cell death). Pretreatment of cells with Taxol and cold storage in the UW solution with 1.5 mM Ca suppressed both bleb formation and LDH release in 48-hr cold-stored cells. Conclusions: Cold storage of hepatocytes leads to reperfusion injury and cell death. This can be suppressed with Taxol and Ca. This suggests that hypothermia induces changes in cellular Ca and a disruption of the microtubules, leading to loss of cell viability. Improved liver preservation may require suppression of Ca-dependent disruption of the cytoskeleton system of liver cells.