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2007 | 10 | 1 |
Tytuł artykułu

Hypothermic storage of equine isolated hepatocytes

Warianty tytułu
Języki publikacji
EN
Abstrakty
EN
The aim of the study was to establish the optimal methods for hypothermic storage of equine isolated hepatocytes. Viability of equine isolated hepatocytes after hypothermic storage was dependent on the type of storage medium as well as on the cell density in the storage suspension and the preservation period. Hepatocytes stored at 4°C in Hanks' Balanced Salt Solution (HBSS) and Williams' Medium E (WE) for 24 h showed very low viability, numerous cell membrane blebs, very low attachment rate (11.9 ± 6.5% and 34.8 ± 19.1%, respectively) and 3-(4,5-dimethylthiazol-2-yl)-2,5- diphenyltetrazolium bromide (MTT) reduction rate (6.4 ± 3.9% and 25.1 ± 14.8%, respectively). In contrast, hepatocytes stored in University of Wisconsin Solution (UW) after 24 h of storage at a density of 12.5 x 106 cells/ml showed high viability (over 70%), typical and intact morphology, high cell attachment rates and MTT reduction. Our findings clearly demonstrate that UW is a good preservation solution for equine isolated hepatocytes. Hepatocytes harvested from slaughterhouse organs can be stored at 4°C in UW at a density of 12.5 x 106 cells/ml for at least 24 h without significant decrease in functional integrity.
Wydawca
-
Rocznik
Tom
10
Numer
1
Opis fizyczny
p.11-18,fig.,ref.
Twórcy
autor
  • Warsaw Agricultural University, Ciszewskiego 8, 02-786 Warsaw, Poland
autor
autor
Bibliografia
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  • Bąkała A, Karlik W, Wiechetek M (2003b) Monolayer cul­ture of equine hepatocytes: conditions for cell attach­ment. J Vet Pharmacol Therap 26 (Suppl. 1): 300.
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  • Donato MT, Castell JV, Gomez-Lechon MJ (1998) The coumarin 7-hydroxylation microassay in living hepatic cells in culture. ATLA 26: 213-223.
  • Gores GJ, Herman B, Lemasters JJ (1990) Plasma mem­brane bleb formation and rupture: a common feature of hepatocellular injury. Hepatology 11: 690-698.
  • Guillouzo A, Guyomard C, Fautrel A, Chesne C (2000) Storage of isolated hepatocytes. In: Berry MN, Edwards AM (eds) The Hepatocyte Review. Kluwer Academic Publishers, pp 125-145.
  • Guyomard C, Chesné C, Meunier B, Fautrel A, Clerc C, Morel F, Rissel M, Campion JP, Guillouzo A (1990) Primary culture of adult rat hepatocytes after 48-hour preservation of the liver with cold UW solution. Hepatol­ogy 12: 1329-1336.
  • Koebe HG, Pahernik SA, Thasler WE, Schildberg FW (1996) Porcine hepatocytes for biohybrid artificial liver devices: a comparison of hypothermic storage techniques. Artif Organs 20: 1181-1190.
  • Li G, Liu Y, Liand J (2003) Isolation and protective effect in UW solution of human hepatocytes during cold storage. International Congress Series 1255: 217-218.
  • Marsh DC, Hjelmhaug JA, Vreugdenhil PK, Kerr JA, Rice MJ, Belzer FO, Southard JH (1991) Hypothermic preservation of hepatocytes. III. Effects of resuspension me­dia on viability after up to 7 days of storage. Hepatology 13: 500-508.
  • Meredith MJ (1988) Rat hepatocytes prepared without col­lagénase: prolonged retention of differentiated charac­ters in culture. Cell Biol Toxicol 4: 405-425.
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  • Nyberg SL, Shatford RA, Pagne WD, Hu WS, Cerra FB (1993) Staining with fluorescein diacetate correlates with hepatocyte function. Biotech Histochem 68: 56-63.
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  • Ostrowska A, Bode DC, Pruss J, Bilir B, Smith GD, Zeislaft S (2000) Investigation of functional and morphological integrity of freshly isolated and cryopreserved human hepatocytes. Cell Tissue Banking 1: 55-68.
  • Pahernik SA, Thasler WE, Mueller-Hoecker J, Schildberg FW, Koebe HG (1996) Hypothermic storage of pig hepatocytes: influence of different storage solutions and cell density. Cryobiology 33: 552-566.
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  • Poullain MG, Fautrel A, Guyomard C, Chesne C, Grislain L, Guillouzo A (1992) Viability and primary culture of rat hepatocytes after hypothermic preservation: the su­periority of the Leibovitz medium over the University of Wisconsin solution for cold storage. Hepatology 15: 97-106.
  • Rissel M, Guiguen K, Meunier B, Joly B, Fautrel A, Guillouzo A (1993) Viability and function of rat and hu­man hepatocytes after hypothermic preservation. Hepa­tology 18: 321A.
  • Sandker GW, Slooff MJH, Groothuis GMM (1992) Drug transport, viability and morphology of isolated rat hepa­tocytes preserved for 24 h in University of Wisconsin sol­ution. Biochem Pharmacol 43: 1479-1485.
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Typ dokumentu
Bibliografia
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