Abstract
Cryopreserved porcine hepatocytes in collagen cultures secreted albumin (up to 98 ± 3 μg ml−1; non-cryopreserved controls: 100 ± 9 μg ml−1) and metabolised diazepam (67.5% ± 7.5% of initially applied diazepam are metabolised; 77.5% ± 10% in non-cryopreserved controls) for up to 14 days after thawing. The cultures resembled the recently developed flat membrane bioreactors. Addition of 5% (v/v) serum did not effect diazepam metabolism. Hepatocytes in collagen-sandwich cultures offer a storable liver support system for potential clinical use.
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Bader A, Knop E, Boeker K, Fruehauf N, Schuettler W, Oldhafer K, Pichlmayer R, Sewing KF (1995) A novel bioreactor design for in vitro reconstruction of in vivo liver characteristics. Artif. Organs 19: 368-374.
Burnell JM, Runge C, Saunders FC, Thomas ED, Volwiler W (1973) Acute hepatic failure treated by cross circulation. Arch. Int. Med. 132: 493-498.
Denis J, Opolon P, Nusinovici V, Granger A, Darnis F (1978) Treatment of encephalopathy during fulminant hepatic failure by haemodialysis with high permeability membrane. Gut 19: 787-793.
Dich J, Vind C, Grunnet N (1987) Long-term culture of hepatocytes: ethanol oxidation and effect of ethanol on enzyme activities and albumin secretion. Alcohol 1: 271-5.
Elsdale T, Bard J (1972) Collagen substrata for studies on cell behaviour. J. Cell. Biol. 54: 626-637.
Diener B, Utesch D, Beer N, Duerk H, Oesch F (1993) A method for the cryopreservation of liver parenchymal cells for studies of xenobiotics. Cryobiology 30: 116-127.
Gerlach J, Encke J, Hole O, Mueller C, Courtney J, Neuhaus P (1994) Hepatocyte culture between three dimensional arranged biomatrix-coated independent artificial capillary systems and sinusoidal endothelial cell co-culture compartments. Int. J. Artif. Organs 17: 301-306.
Gerlach J, Kloppel K, Schauwecker HH, Tauber R, Mueller C, Bucherl ES (1989) Use of hepatocytes in adhesion and suspension cultures for liver support bioreactors. Int. J. Artif. Organs 12: 788-792.
Guguen-Guillouzo G, Clement B, Buffet G, Beaumont C, Morel-Chany E, Glaise D, Guillouzo A (1983) Maintenance and reversibility of active albumin secretion by adult rat hepatocytes cocultured with another liver epithelial cell type. Exp. Cell Res. 143: 47-54.
Hoogenboom LA, Pastoor FJ, Clous WE, Hesse SE, Kuiper HA (1989) Use of porcine hepatocytes for biotransformation studies of veterinary drugs. Xenobiotica 19: 1207-1219.
Hu MY, Cipolle M, Sielaff T, Lovdahl MJ, Mann HJ, Remmel RP, Cerra FB (1995) Effects of hepatocyte growth factor on viability and biotransformation functions on hepatocytes in gel entrapped and monolayer culture. Crit. Care Med. 23: 1237-1242.
Koebe HG, Daehnhardt C, Mueller-Hoecker J, Wagner H, Schildberg FW (1996) Cryopreservation of porcine hepatocyte cultures Cryobiology 33: 127-141.
Koide N, Shinji T, Tanabe T, Asano K, Kawaguchi M, Sakaguchi K, Koide Y, Mori M, Tsuji T (1989) Continued high albumin production by multicellular spheroids of adult rat hepatocytes in the presence of liver derived proteoglycans. Biochem. Biophys. Res. Commun. 161: 385-391.
Lazar A, Mann HJ, Remmel RP, Shatford RA, Cerra FB, Hu WS (1995) Extended liver-specific functions of porcine hepatocyte spheroids entrapped in collagen gel. In Vitro Cell Dev. Biol. Anim. 31: 340-346.
Mullen KD, Jones EA (1996) Natural benzodiazepines and hepatic encephalopathy. Semin. Liv. Dis. 16: 255-264.
O'Grady JG, Gimson AES, O'Brien CJ, Pucknell A, Hughes RD, Williams R (1988) Controlled trials of charcoal haemoperfusion and prognostic factors in fulminant hepatic failure. Gastroenterology 94: 1186-1192.
Parbhoo SP, James IM, Ajdukiewicz A, Xanalatos C, Kennedy J, Chalstrey LJ, Brock PJ, Sayer P, Sherlock S (1971) Extracorporeal pig-liver perfusion in treatment of hepatic coma due to fulminant hepatitis. Lancet 1: 659-665.
Redeker AG, Yamahiro HS (1973) Controlled trial of exchangetransfusion therapy in fulminant hepatitis. Lancet 1: 3-6.
Stange J, Mitzner S (1996) Hepatocyte encapsulation-initial intentions and new aspects for its use in bioartificial liver support. Int. J. Artif. Organs 19: 45-48.
Strom SC, Jirtle RL, Jones RS, Novicki DL, Rosenberg MR, Novotny A, Irons G, McLain JR, Michalopoulos G (1982) Isolation, culture, and transplantation of human hepatocytes. J. Natl. Cancer Inst. 68: 771-778.
Unseld E, Krishna DR, Fischer C, Klotz U (1989) Detection of desmethyl diazepam and diazepam in brain of different species and plants. Biochem. Pharmacol. 38: 2473-2478.
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Karim, N., Allmeling, C., Hengstler, JG. et al. Diazepam metabolism and albumin secretion of porcine hepatocytes in collagen-sandwich after cryopreservation. Biotechnology Letters 22, 1647–1652 (2000). https://doi.org/10.1023/A:1005685912195
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DOI: https://doi.org/10.1023/A:1005685912195