2011, Number 2
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Ann Hepatol 2011; 10 (2)
Hypothermic preservation of rat liver microorgans (LMOs) in bes-gluconate solution. Protective effects of polyethyleneglycol (PEG) on total water content and functional viability
Mandolino C, Pizarro MD, Quintana AB, Rodríguez JV, Mamprin ME
Language: English
References: 29
Page: 196-206
PDF size: 360.31 Kb.
ABSTRACT
We have reported of an alternative solution to preserve hepatocytes that have three key components:
gluconate, sucrose and an aminosulfonic acid (BGS solution). In order to extend the use of this solution to
organs as the liver, we evaluate the effect of the addition of PEG of 8, 20 and 35 kDa to BG Solution on the
total water content and functional viability of rat liver microorgans (LMOs). LMOs were preserved (48 h 0
ºC) in the following solutions: ViaSpan
®; BGS; BG plus 4% PEG 8000 (BG8); BG plus 4% PEG 20.000 (BG20) and
BG plus 4% PEG 35.000 (BG35). LDH Release and Total Water Content showed a marked increase in LMOs
preserved in BGS. This indicates that, in the absence of PEG, the tissue showed important cell membrane
integrity deterioration and was incapable of regulating cell volume. After the preservation period, all
groups were reoxygenated (120 min, 37
ºC, KHR) and Total Water Content, Glycogen Content and Oxygen
Consumption were determined. After 120 min LMOs preserved in BG35 showed values of Oxygen Consumption
similar to controls. On the other hand, LMOs preserved in BG8, BG20 and ViaSpan
® showed oxygen
consumption rates and glycogen content significantly smaller than controls. In conclusion, BG35 was
the most effective preservation solution to protect LMOs against cold preservation injury due to ischemia
and reoxygenation. It is a good alternative to ViaSpan
® because of its higher buffer capacity, its best indexes
of respiration activity and for being considerably less expensive.
REFERENCES
Kamada N, Calne RY. A surgical experience with five hundred thirty liver transplants in the rat. Surgery 1983; 93: 64-9.
Sumimoto R, Lindell S, Gambiez L, Southard JH, Belzer FO. The effect of pretransplant flushout of the liver with a solution containing albumin on survival and organ function. Transplant Proc 1993; 25: 3001-3.
D’ Alessandro AM, Southard JH, Kalayoglu M, Belzer FO. Comparison of cold stotage and perfusion of dog livers on function of tissue slices. Cryobiology 1986; 23: 161-7.
Vreugdenhil PK, Marsh DC, Southard JH. Comparison of isolated hepatocytes and tissue slices for study of liver hypothermic preservation/reperfusion injury. Cryobiology 1996; 33: 430-5.
Mamprin ME, Petrocelli S, Guibert EE, Rodríguez JV. A novel Bes-Gluconate-Sucrose (BGS) solution for cold storage of isolated hepatocytes. CryoLetters 2008; 29: 121-33.
Yoshida H, Okuno H, Kamoto T, Habuchi T, Toda Y, Hasegawa S, Nakamura T, et al. Transplantation 2002; 74: 1231-6.
Kravchenko LP, Petrenko AY, Somov A, Grischenko V, Fuller BJ. Respiratory activity of isolated rat hepatocytes following cold storage and subsequent rewarming: a comparison of sucrose-based and University of Wisconsin solution. Cryobiology 2001; 42: 218-21.
Rees D. The preservation of cellular function in isolated mammalian tissues/organ preparations. Proc Physiol Soc N Z 1988; 8: 21.
Kasper H, Konze E, Kutinová Canová N, Dienes H, Dries V. Cryopreservation of precision cut tissue slices (PCTS): Investigation of morphology and reactivity. Exp Toxicol Pathol 2010. In press.
Hosgood SA, Nicholson ML. Hydrogen sulphide ameliorates ischaemia-reperfusion injury in an experimental model of non-heart-beating donor kidney transplantation. Br J Surg 2010; 2: 202-20.
Martin DR, Scott DF, Downes GL, Belzer FO. Primary cause of unsuccessful liver and heart transplantation: cold sensitivity of the ATPase system. Ann Surg 1972; 175: 111- 17.
Southard JH, Belzer FO. Control of canine Kidney cortex slice volume and ion distribution at hypothermia by impermeable anions. Cryobiology 1980; 17: 540-8.
Howden B, Jablonski P, Thomas A, Walls K, Biguzas M, Scott DF, Grossman H, et al. Liver preservation with UW solution. I. Evidence that hydroxyethyl starch is not essential. Transplantation 1990; 49: 869-72.
van der Plaats A, ‘t Hart NA, Morariu AM, Verkerke GJ, Leuvenink HG, Ploeg RJ, Rakhorst G. Effect of University of Wisconsin organ-preservation solution on haemorheology. Transpl Int 2004; 17: 227-33.
Giraud S, Claire B, Eugene M, Debre P, Richard F, Barrou B. A new preservation solution increases islet yield and reduces graft immunogenicity in pancreatic islet transplantation. Transplantation 2007; 83: 1397-400.
Faure JP, Hauet T, Han Z, Goujon JM, Petit I, Mauco G, Eugene M, et al. Polyethylene glycol reduces early and longterm cold ischemia-reperfusion and renal medulla injury. J Pharmacol Exp Ther 2002; 302: 861-70.
Neuzillet Y, Giraud S, Lagorce L, Eugene M, Debre P, Richard F, Barrou B. Effects of the Molecular Weight of PEG Molecules (8, 20, 35 KDa) on Cell Function and Allograft Survival Prolongation in Pancreatic Islets Transplantation. Transplantation proceedings 2006; 38: 2354-5.
Mamprin ME, Guibert EE, Rodríguez JV. Glutathione content during the rinsing and rewarming process of rat hepatocytes preserved in University of Wisconsin solution. Cryobiology; 40: 270-276.
Sumimoto R, Jamieson NV, Wake K, Kamada N. 24-hour rat liver preservation using UW solution and some simplified variants. Transplantation 1989; 48: 1-5.
Roe JH, Epstein J, Goldstein N. A photometric method for the determination of inulin in plasma and urine. J Biol Chem 1949; 839-45.
Bergmeyer HU, Bernt E. Lactate deshydrogenase. In: Bergmeyer, HU (eds.). “Methods in Enzymatic Analyse”. Vol. 2. 2nd. Ed. Verlag Chemie, Weinheim/Bergsts; p. 1407-10.
Carr RS, Neff JM. Quantitative semi-automated enzimatic assay for tissue glycogen. Comparative Biochemistry Physiology 1984; 3: 447-9.
Robinson J, Cooper J. Method of determining oxygen concentrations in biological media, suitable for calibration of oxygen electrode. Anal biochemistry 1970; 33: 390-9.
De Kanter R, Monshouwer M, Meijer D, Groothuis G. Precision- cut organ slices a tool to study toxicity and metabolism of xenobiotics with special reference to non-hepatic tissue. Current Drug Metabolism 2002; 3: 39-59.
Olinga P, Groen K, Hof H, De Kanter R, Koster HJ, Leeman WR, Rutten AA, et al. Comparison of five incubation systems for rat liver slices using functional and viability parameters. J Pharmacol Toxicol Methods 1997; 2: 59-69.
Belzer FO, Southard JH. Principle of solid-organ preservation by cold storage. Transplantation 1988; 45: 676.
Lagadic-Gossman D, Rissel M, Le Bot MA, Guillouzo A. Toxic effects of tacrine primary hepatocytes and liver epithelial cells in culture. Cell Biol. Toxicol 1998; 5: 361-73.
Kantrow S, Taylor D, Caraway M, Piantadosi C. Oxidative metabolism in rat hepatocytes and mitochondria during sepsis. Arch Biochem Biophys 1997; 345: 278-88.
Kim S, Belzer FO, Southard J.H. Loss of mitochondrial respiratory function and its suppression during cold ischemic preservation of rat livers with university of Wisconsin solution. Hepatology 1992: 16: 742-8.