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Biochem J. Mar 15, 1997; 322(Pt 3): 693–699.
PMCID: PMC1218244

Rapid decrease in cellular sodium and chloride content during cold incubation of cultured liver endothelial cells and hepatocytes.


Hypothermia, as used for organ preservation in transplantation medicine, is generally supposed to lead to an intracellular accumulation of sodium, and subsequently of chloride, via inhibition of the Na+/K+-ATPase. However, on studying the cellular sodium concentration of cultured liver endothelial cells using fluorescence microscopy, we found a 55% decrease in the cellular sodium concentration after 30 min of cold incubation in University of Wisconsin (UW) solution. To confirm this surprising result, we set up a capillary electrophoresis method that allowed us to determine the cellular contents of inorganic cations and of inorganic anions. Using this method we measured a decrease in the cellular sodium content from 104+/-11 to 55+/-4 nmol/mg of protein, accompanied by a decrease in the chloride content from 71+/-9 to 25+/-5 nmol/mg of protein, after 30 min of cold incubation in UW solution. When the endothelial cells were incubated in cold Krebs-Henseleit buffer or in cold cell culture medium instead of UW solution, similar early decreases in cellular sodium and chloride contents were observed, thus excluding the possibility of the decreases being dependent on the preservation solution used. Furthermore, experiments with cultured rat hepatocytes yielded a similar decrease in sodium content during initiation of cold incubation in UW solution, so the decrease does not appear to be cell-specific either. These results suggest that, contrary to current opinion, sodium efflux predominates over sodium influx during the early phase of cold incubation of cells.

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Selected References

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  • Lemasters JJ, Bunzendahl H, Thurman RG. Reperfusion injury to donor livers stored for transplantation. Liver Transpl Surg. 1995 Mar;1(2):124–138. [PubMed]
  • Blankensteijn JD, Terpstra OT. Liver preservation: the past and the future. Hepatology. 1991 Jun;13(6):1235–1250. [PubMed]
  • Clavien PA, Harvey PR, Strasberg SM. Preservation and reperfusion injuries in liver allografts. An overview and synthesis of current studies. Transplantation. 1992 May;53(5):957–978. [PubMed]
  • Kruuv J, Glofcheski DJ, Lepock JR. Evidence for two modes of hypothermia damage in five cell lines. Cryobiology. 1995 Apr;32(2):182–190. [PubMed]
  • Hansen TN, Dawson PE, Brockbank KG. Effects of hypothermia upon endothelial cells: mechanisms and clinical importance. Cryobiology. 1994 Feb;31(1):101–106. [PubMed]
  • Hochachka PW. Defense strategies against hypoxia and hypothermia. Science. 1986 Jan 17;231(4735):234–241. [PubMed]
  • Belzer FO, Southard JH. Principles of solid-organ preservation by cold storage. Transplantation. 1988 Apr;45(4):673–676. [PubMed]
  • Larsen T, Solberg S, Johansen R, Jørgensen L. Effect of cooling on the intracellular concentrations of Na+, K+ and Cl- in cultured human endothelial cells. Scand J Clin Lab Invest. 1988 Oct;48(6):565–571. [PubMed]
  • Caldwell-Kenkel JC, Currin RT, Tanaka Y, Thurman RG, Lemasters JJ. Kupffer cell activation and endothelial cell damage after storage of rat livers: effects of reperfusion. Hepatology. 1991 Jan;13(1):83–95. [PubMed]
  • McKeown CM, Edwards V, Phillips MJ, Harvey PR, Petrunka CN, Strasberg SM. Sinusoidal lining cell damage: the critical injury in cold preservation of liver allografts in the rat. Transplantation. 1988 Aug;46(2):178–191. [PubMed]
  • Rauen U, Hanssen M, Lauchart W, Becker HD, de Groot H. Energy-dependent injury to cultured sinusoidal endothelial cells of the rat liver in UW solution. Transplantation. 1993 Mar;55(3):469–473. [PubMed]
  • Rauen U, Hintz K, Hanssen M, Lauchart W, Becker HD, de Groot H. Injury to cultured liver endothelial cells during cold preservation: energy-dependent versus energy-deficiency injury. Transpl Int. 1993;6(4):218–222. [PubMed]
  • de Groot H, Brecht M. Reoxygenation injury in rat hepatocytes: mediation by O2/H2O2 liberated by sources other than xanthine oxidase. Biol Chem Hoppe Seyler. 1991 Jan;372(1):35–41. [PubMed]
  • Harootunian AT, Kao JP, Eckert BK, Tsien RY. Fluorescence ratio imaging of cytosolic free Na+ in individual fibroblasts and lymphocytes. J Biol Chem. 1989 Nov 15;264(32):19458–19467. [PubMed]
  • Kawanishi T, Nieminen AL, Herman B, Lemasters JJ. Suppression of Ca2+ oscillations in cultured rat hepatocytes by chemical hypoxia. J Biol Chem. 1991 Oct 25;266(30):20062–20069. [PubMed]
  • Jung DW, Apel LM, Brierley GP. Transmembrane gradients of free Na+ in isolated heart mitochondria estimated using a fluorescent probe. Am J Physiol. 1992 Apr;262(4 Pt 1):C1047–C1055. [PubMed]
  • O'Neill WC, Klein JD. Regulation of vascular endothelial cell volume by Na-K-2Cl cotransport. Am J Physiol. 1992 Feb;262(2 Pt 1):C436–C444. [PubMed]
  • Navas JP, Anderson W, Marsh JD. Hypothermia increases calcium content of hypoxic myocytes. Am J Physiol. 1990 Aug;259(2 Pt 2):H333–H339. [PubMed]
  • Brock TA, Brugnara C, Canessa M, Gimbrone MA., Jr Bradykinin and vasopressin stimulate Na+-K+-Cl- cotransport in cultured endothelial cells. Am J Physiol. 1986 Jun;250(6 Pt 1):C888–C895. [PubMed]
  • Minta A, Tsien RY. Fluorescent indicators for cytosolic sodium. J Biol Chem. 1989 Nov 15;264(32):19449–19457. [PubMed]
  • Levi AJ, Lee CO, Brooksby P. Properties of the fluorescent sodium indicator "SBFI" in rat and rabbit cardiac myocytes. J Cardiovasc Electrophysiol. 1994 Mar;5(3):241–257. [PubMed]
  • Negulescu PA, Machen TE. Intracellular ion activities and membrane transport in parietal cells measured with fluorescent dyes. Methods Enzymol. 1990;192:38–81. [PubMed]
  • Churchill TA, Fuller BJ. Effects of sodium and potassium channel antagonists on the energy metabolism of rat liver during cold storage. Transplantation. 1995 Mar 27;59(6):904–907. [PubMed]
  • Kimzey SL, Willis JS. Temperature adaptation of active sodium-potassium transport and of passive permeability in erythrocytes of ground squirrels. J Gen Physiol. 1971 Dec;58(6):634–649. [PMC free article] [PubMed]
  • Murphy E, Aiton JF, Horres CR, Lieberman M. Calcium elevation in cultured heart cells: its role in cell injury. Am J Physiol. 1983 Nov;245(5 Pt 1):C316–C321. [PubMed]
  • Schmitt M, Saulnier F, Malhautier L, Linden G. Effect of temperature on the salt balance of milk studied by capillary ion electrophoresis. J Chromatogr. 1993 Jun 25;640(1-2):419–424. [PubMed]
  • Levinson C. Regulatory volume increase in Ehrlich ascites tumor cells is mediated by the 1Na:1K:2Cl cotransport system. J Membr Biol. 1992 Mar;126(3):277–284. [PubMed]
  • Verkman AS. Development and biological applications of chloride-sensitive fluorescent indicators. Am J Physiol. 1990 Sep;259(3 Pt 1):C375–C388. [PubMed]
  • Chao AC, Dix JA, Sellers MC, Verkman AS. Fluorescence measurement of chloride transport in monolayer cultured cells. Mechanisms of chloride transport in fibroblasts. Biophys J. 1989 Dec;56(6):1071–1081. [PMC free article] [PubMed]
  • Jensen BS, Jessen F, Hoffmann EK. Na+, K+, Cl- cotransport and its regulation in Ehrlich ascites tumor cells. Ca2+/calmodulin and protein kinase C dependent pathways. J Membr Biol. 1993 Feb;131(3):161–178. [PubMed]
  • Hall SR, Sigee DC, Beesley JE. Scanning X-ray microanalysis of microcarrier cultured endothelial cells: elemental changes during the transition to confluency and the effect of ionophore A23187. Scanning Microsc. 1992 Sep;6(3):753–763. [PubMed]
  • Quang C, Khaledi MG. Prediction and optimization of the separation of metal cations by capillary electrophoresis with indirect UV detection. J Chromatogr A. 1994 Jan 28;659(2):459–466. [PubMed]
  • Knerr SM, Lieberman M. Ion transport during hypothermia in cultured heart cells: implications for protection of the immature myocardium. J Mol Cell Cardiol. 1993 Mar;25(3):277–288. [PubMed]
  • Crowe JH, Hoekstra FA, Crowe LM, Anchordoguy TJ, Drobnis E. Lipid phase transitions measured in intact cells with Fourier transform infrared spectroscopy. Cryobiology. 1989 Feb;26(1):76–84. [PubMed]
  • Rauen U, Elling B, Gizewski ER, Korth HG, Sustmann R, de Groot H. Involvement of reactive oxygen species in the preservation injury to cultured liver endothelial cells. Free Radic Biol Med. 1997;22(1-2):17–24. [PubMed]

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