The influence of medium composition on the maintenance of cytochrome P-450, glutathione content and urea synthesis: a comparison of rat and sheep primary hepatocyte cultures

Watts, Patricia; Smith, Mark D.; Edwards, Ian; Zammit, Victor A.; Brown, Vivienne; Grant, Helen

doi:10.1016/0168-8278(95)80069-7

Cited by 27 publications

(10 citation statements)

References 23 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…3). This concurred with values from serum-containing cultures of rat, mouse, hamster and human hepatocytes3055. The labile nature of total P450 may be explained by observed down-regulation of CYP450 mRNA transcripts triggered during the collagenase isolation process56.…”

Section: Discussionsupporting

confidence: 78%

See 1 more Smart Citation

Profiling the Impact of Medium Formulation on Morphology and Functionality of Primary Hepatocytes in vitro

Nelson

Treskes

Howie

et al. 2013

Sci Rep

View full text Add to dashboard Cite

The characterization of fully-defined in vitro hepatic culture systems requires testing of functional and morphological variables to obtain the optimal trophic support, particularly for cell therapeutics including bioartificial liver systems (BALs). Using serum-free fully-defined culture medium formulations, we measured synthetic, detoxification and metabolic variables of primary porcine hepatocytes (PPHs) - integrated these datasets using a defined scoring system and correlated this hepatocyte biological activity index (HBAI) with morphological parameters. Hepatic-specific functions exceeded those of both primary human hepatocytes (PHHs) and HepaRG cells, whilst retaining biotransformation potential and in vivo-like ultrastructural morphology, suggesting PPHs as a potential surrogate for PHHs in various biotech applications. The HBAI permits assessment of global functional capacity allowing the rational choice of optimal trophic support for a defined operational task (including BALs, hepatocellular transplantation, and cytochrome P450 (CYP450) drug metabolism studies), mitigates risk associated with sub-optimal culture systems, and reduces time and cost of research and therapeutic applications.

show abstract

Section: Discussionsupporting

confidence: 78%

“…Previous studies have shown that the type of medium formulation modulates morphology and function of rat and sheep hepatocyte cultures2930, as well as porcine hepatocytes on biomatrix-coated culture dishes with or without serum131.…”

mentioning

confidence: 99%

Profiling the Impact of Medium Formulation on Morphology and Functionality of Primary Hepatocytes in vitro

Nelson

Treskes

Howie

et al. 2013

Sci Rep

View full text Add to dashboard Cite

show abstract

“…Total cytochrome P450 content was measured in the freshly isolated cells by a modification of the original method of Omura and Sato (1964) as detailed by Watts et al. (1995).…”

Section: Methodsmentioning

confidence: 99%

Metabolism of isometamidium in hepatocytes isolated from control and inducer‐treated rats

Boibessot

Tettey

Skellern

et al. 2006

Vet Pharm & Therapeutics

View full text Add to dashboard Cite

Little is known about the metabolism and mechanism of action of the trypanocide, isometamidium (ISM), the major drug used for prophylaxis of trypanosomiasis. We have investigated its metabolism and distribution in isolated rat hepatocytes using liquid chromatography-mass spectrometry and confocal laser scanning microscopy (CLSM). Two putative metabolites were formed, which were proposed to be a mono-acetyl derivative and an oxidized metabolite (SII). This is the first demonstration of the hepatic metabolism of ISM, as previous in vivo studies were hampered by dose-limiting toxicity and insensitive analytical methods. The intrinsic fluorescence of the drug enabled its intracellular uptake to be followed by CLSM. It is taken up rapidly into the nucleolus, nuclear membrane and endoplasmic reticulum within 5 min, and retained in the nucleus for at least 24 h. Persistent binding of ISM to cellular macromolecules may contribute to its prophylactic effect in vivo. Pretreatment of rats with 3-methylcholanthrene, phenobarbitone (PB) or the widely used pyrethroid pesticide, deltamethrin, resulted in an increase in metabolism of ISM to the proposed SII after 1 h incubation with hepatocytes. 3-methylcholanthrene was the most potent inducer, causing a maximal 19.5-fold induction of SII formation after exposure of hepatocytes to ISM for 1 h compared with formation by control hepatocytes. In comparison, at the 1 h timepoint deltamethrin pre-treatment caused a 10.2-fold induction, and PB only 8.2 fold.

show abstract

“…Measurements of cytochromes P450 and b 5 content were carried out as previously described (Watts et al 1995). The absorbance of CO-treated cell homogenates was compared with that of COtreated and sodium dithionite reduced homogenates between wavelengths of 400 and 500 nm.…”

Section: Cytochromes P450 and Bmentioning

confidence: 99%

Metabolism of troglitazone in hepatocytes isolated from experimentally induced diabetic rats

Meechan

Henderson

Bates

et al. 2006

Journal of Pharmacy and Pharmacology

View full text Add to dashboard Cite

Troglitazone (TGZ), the prototype 2,4-thiazolidinedione antidiabetic agent, is associated with hepatotoxicity in patients with Type 2 diabetes. Although the mechanism of toxicity has not been established, alterations in the clearance of TGZ from in-vitro hepatocyte cultures through metabolic conjugation reactions are believed to modulate the toxicity of the compound. In this study, the metabolism of TGZ in freshly isolated hepatocytes from the fat-fed streptozotocin-treated rat model of Type 2 diabetes is described. Biochemical parameters such as cellular reduced glutathione content, content of cytochromes P450 and b5, and the expression of glutathione-S-transferase alpha (subunits Ya and Yc2) were not affected by the induced diabetes. TGZ was metabolized primarily to a sulfonate, a quinone and a glucuronide in both control and experimentally diabetic animals. However, metabolism after induction of diabetes was characterized by a moderate increase in sulfation, a decrease in the elimination half-life of TGZ and the absence of the minor metabolites of TGZ, notably the glutathione adduct of the putative reactive intermediate (m/z = 747 (M + H)+; m/z = 745 (M - H)-).

show abstract

The influence of medium composition on the maintenance of cytochrome P-450, glutathione content and urea synthesis: a comparison of rat and sheep primary hepatocyte cultures

Cited by 27 publications

References 23 publications

Profiling the Impact of Medium Formulation on Morphology and Functionality of Primary Hepatocytes in vitro

Profiling the Impact of Medium Formulation on Morphology and Functionality of Primary Hepatocytes in vitro

Metabolism of isometamidium in hepatocytes isolated from control and inducer‐treated rats

Metabolism of troglitazone in hepatocytes isolated from experimentally induced diabetic rats

Contact Info

Product

Resources

About