Phase I and II metabolism and carbohydrate metabolism in cultured cryopreserved porcine hepatocytes

Loven, Anne-Dorthe Bonde; Olsen, Anette Kristensen; Friis, Christian; Andersen, Birgitte

doi:10.1016/j.cbi.2005.03.040

Cited by 14 publications

(6 citation statements)

References 24 publications

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“…As shown by Loven et al,31 phase I and II enzyme activities in cryopreserved porcine hepatocytes are detectable, but exhibit lower levels compared to freshly isolated cells. Hengstler et al3 found low basal ethoxyresorufin‐O‐deethylase activity in cultured cryopreserved hepatocytes.…”

Section: Discussionmentioning

confidence: 80%

Cryopreservation of primary human hepatocytes: The benefit of trehalose as an additional cryoprotective agent

et al. 2006

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Problems with the limited availability of human hepatocytes for cell transplantation may be overcome by efficient cryopreservation techniques and formation of appropriate cell banking. In this study we investigated the effect of the disaccharide trehalose on the cryopreservation of human hepatocytes. For analysis, liver cells were frozen in culture medium containing 10% dimethyl sulfoxide (DMSO) that was supplemented with varying concentrations of trehalose. During the postthawing culture period, viability, plating efficiency, total protein, cell proliferation, enzyme leakage, albumin and urea formation, as well as phase I and II metabolism were analyzed. In the pilot study, among the concentrations investigated, 0.2 M trehalose showed the best overall outcome. Compared to the use of DMSO alone, we found significant improvement in postthaw cell viability (62.9 Ϯ 13 vs. 46.9 Ϯ 11%, P Ͻ 0.01) and plating efficiency (41.5 Ϯ 18 vs. 17.6 Ϯ 13%, P Ͻ 0.01) in the trehalose group. The use of trehalose as an additive for cryopreserving human hepatocytes resulted in a significantly increased total protein level in the attached cells, higher secretion of albumin and a lower aspartate aminotransferase (AST) level after thawing. In conclusion, the use of trehalose as cryoprotective agent significantly improves the outcome of human hepatocyte cryopreservation. Liver Transpl 13:38-45, 2007.

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Section: Discussionmentioning

confidence: 80%

Cryopreservation of primary human hepatocytes: The benefit of trehalose as an additional cryoprotective agent

et al. 2006

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“…Basal and glucagon induced-glycogenolysis were measured as glucose released into the buffer, using the glucose oxidase kit (Sigma) as described previously (Loven et al, 2005). …”

Section: Methodsmentioning

confidence: 99%

S-Adenosylmethionine regulates connexins sub-types expressed by hepatocytes

Yamaji

Droggiti

et al. 2011

European Journal of Cell Biology

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Intercellular communication via GAP Junctions plays an important role in tissue homeostasis, apoptosis, carcinogenesis, cell proliferation and differentiation. Hepatocyte connexins (Cx) 26 and 32 levels are decreased during the de-differentiation process of primary hepatocytes in culture, a situation that is also characterised by a decrease in S-Adenosylmethionine (SAMe) levels. In this current study, we show that SAMe supplementation in cultured hepatocytes every 12h, leads to an up-regulation of Cx26 and 32 mRNA and protein levels and blocks culture-induced Cx43 expression, although it failed to increase Cx 26 and 32 membrane localization and GAP junction intracellular communication. SAMe reduced nuclear β-catenin accumulation, which is known to stimulate the TCF/LEF-dependent gene transcription of Cx43. Moreover SAMe-induced reduction in Cx43 and β-catenin- was prevented by the proteasome inhibitor MG132, and was not mediated by GSK3 activity. SAMe, and its metabolite 5′-Methyltioadenosine (MTA) increased Cx26 mRNA in a process partially mediated by Adenosine A2A receptors but independent of PKA. Finally livers from MAT1A knockout mice, characterized by low hepatic SAMe levels, express higher Cx43 and lower Cx26 and 32 protein levels than control mice. These results suggest that SAMe maintains a characteristic expression pattern of the different Cxs in hepatocytes by differentially regulating their levels.

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“…In many reports, cryopreservation techniques of primary hepatocytes have been developed using various cryoprotectants and/or programmable cell freezers, but none has conclusively given satisfactory results (Darr & Hubel 2001, Baust et al 2002, Graaf et al 2002, Loven et al 2005.…”

Section: Resultsmentioning

confidence: 99%

Immobilization of Cryopreserved Primary Rat Hepatocytes for the Development of a Bioartificial Liver System

Son

Kim

et al. 2006

Biotechnol Lett

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Primary rat hepatocytes were cryopreserved in hormonally-defined medium (HDM) containing 40% (v/v) fetal bovine serum (FBS) and 10% (v/v) dimethyl sulfoxide (DMSO) in liquid N2 for 6 months. After thawing, the cells were immobilized using 2% (w/v) alginate and 0.5% (w/v) chitosan solutions. The capacities of ammonia removal and urea synthesis of the immobilized-thawed hepatocytes were similar to those of immobilized hepatocytes without cryopreservation. This result shows that immobilized hepatocytes after cryopreservation are useful for the development of a bioartificial liver system.

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Phase I and II metabolism and carbohydrate metabolism in cultured cryopreserved porcine hepatocytes

Cited by 14 publications

References 24 publications

Cryopreservation of primary human hepatocytes: The benefit of trehalose as an additional cryoprotective agent

Cryopreservation of primary human hepatocytes: The benefit of trehalose as an additional cryoprotective agent

S-Adenosylmethionine regulates connexins sub-types expressed by hepatocytes

Immobilization of Cryopreserved Primary Rat Hepatocytes for the Development of a Bioartificial Liver System

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