Optimizing preincubation conditions for precision-cut rat kidney and liver tissue slices: effect of culture media and antioxidants

Obatomi, David K.; Brant, Stephen; Anthonypillai, V.; Early, D.A.; Bach, Peter H.

doi:10.1016/s0887-2333(98)00055-1

Cited by 18 publications

(15 citation statements)

References 37 publications

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“…The damaged cells result during Van Midwoud et al: Hydrogel Embedding of Precision-Cut Liver Slices preparation of the tissue slices, and are only present in the outer layers of the slice. The energy status of the cells also recovers during pre-incubation, as the intracellular adenosine triphosphate (ATP) is restored to the value of fresh tissue after being reduced during the slicing procedure at 48C (De Kanter et al, 2002;Obatomi et al, 1998).…”

Section: Incubation Of Precision-cut Liver Slicesmentioning

confidence: 99%

Hydrogel embedding of precision‐cut liver slices in a microfluidic device improves drug metabolic activity

Midwoud

Merema

Verweij

et al. 2011

Biotech & Bioengineering

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A microfluidic-based biochip made of poly-(dimethylsiloxane) was recently reported for the first time by us for the incubation of precision-cut liver slices (PCLS). In this system, PCLS are continuously exposed to flow, to keep the incubation environment stable over time. Slice behavior in the biochip was compared with that of slices incubated in well plates, and verified for 24 h. The goal of the present study was to extend this incubation time. The viability and metabolic activity of precision-cut rat liver slices cultured in our novel microflow system was examined for 72 h. Slices were incubated for 1, 24, 48, and 72 h, and tested for viability (enzyme leakage (lactate dehydrogenase)) and metabolic activity (7-hydroxycoumarin (phase II) and 7-ethoxycoumarin (phase I and II)). Results show that liver slices retained a higher viability in the biochip when embedded in a hydrogel (Matrigel) over 72 h. This embedding prevented the slices from attaching to the upper polycarbonate surface in the microchamber, which occurred during prolonged (>24 h) incubation in the absence of hydrogel. Phase II metabolism was completely retained in hydrogel-embedded slices when medium supplemented with dexamethasone, insulin, and calf serum was used. However, phase I metabolism was significantly decreased with respect to the initial values in gel-embedded slices with medium supplements. Slices were still able to produce phase I metabolites after 72 h, but at only about ∼10% of the initial value. The same decrease in metabolic rate was observed in slices incubated in well plates, indicating that this decrease is due to the slices and medium rather than the incubation system. In conclusion, the biochip model was significantly improved by embedding slices in Matrigel and using proper medium supplements. This is important for in vitro testing of drug metabolism, drug-drug interactions, and (chronic) toxicity.

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Section: Incubation Of Precision-cut Liver Slicesmentioning

confidence: 99%

Hydrogel embedding of precision‐cut liver slices in a microfluidic device improves drug metabolic activity

Midwoud

Merema

Verweij

et al. 2011

Biotech & Bioengineering

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“…This level of DMSO was nontoxic. The appropriate concentrations of chemical agents used were extrapolated from previously established studies in other in vitro systems (Allman et al 1967;Ishii and Bracht 1986;Wolfgang et al 1989;Toskulkao et al 1994;Obatomi et al 1998a;Schnellmann and Williams 1998).…”

Section: Preparation Of Atractyloside and Tested Compoundsmentioning

confidence: 99%

“…A number of chemical agents used in in vitro and in vivo studies have provided some possibility of therapeutic intervention. We have recently shown that some antioxidants have protective eects on atractyloside-induced toxicity in renal cortical slices (Obatomi et al 1998a). Other workers using pure mitochondria homogenate have also demonstrated that adenosine 5'-diphosphate (ADP), a mitochondrial energy modulator exerts some protective eect on atractyloside toxicity (Allman et al 1967).…”

Section: Introductionmentioning

confidence: 99%

“…The following studies investigated the protective potential of four candidate compounds (adenine nucleotide, stevioside, calpain inhibitor I and probenecid) in renal cortical slices as an extension to an earlier work (Obatomi et al 1998a). The concentration of the various compounds used was based on previous ®ndings (Allman et al 1967;Ishii and Bracht 1986;Wolfgang et al 1989;Toskulkao et al 1994;Obatomi et al 1998a;Schnellmann and Williams 1998).…”

Section: Introductionmentioning

confidence: 99%

“…The concentration of the various compounds used was based on previous ®ndings (Allman et al 1967;Ishii and Bracht 1986;Wolfgang et al 1989;Toskulkao et al 1994;Obatomi et al 1998a;Schnellmann and Williams 1998). The renal cortical slice was chosen as the in vitro kidney model because it contains a population of cells that are selectively targeted by atractyloside and maintained in their normal in vivo architecture (Obatomi et al 1998a(Obatomi et al , 1998b. The parameters selected for the present studies were based on previous experience with this in vitro system and because of their critical roles in reversible and irreversible cell damage.…”

Section: Introductionmentioning

confidence: 99%

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Adenine nucleotide and calpain inhibitor I protect against atractyloside-induced toxicity in rat renal cortical slices in vitro

Obatomi

Blackburn

Bach³

2001

Archives of Toxicology

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Atractyloside is a compound with a documented nephrotoxicity. It induces renal tubular necrosis at high doses and apoptosis at lower doses. This study investigates the potential protective effect of some chemical agents against atractyloside-induced nephrotoxicity in vitro using the precision-cut rat renal cortical slices obtained from kidneys of Wistar rats. For co-incubation experiments, slices were incubated for 3 h at 37 degrees C on a rocker platform with various chemical agents: ADP (5 mM), calpain inhibitor I (CPI, 1 mM), stevioside (STV, 2.5 mM) or probenecid (PRB, 2.5 mM) in the presence or absence of atractyloside (2 mM). For pre-incubation experiments, slices were incubated with the same chemical agents for 1 h before exposure to atractyloside. The nephrotoxic effects of atractyloside (2 mM) alone were manifested in several ways: by a marked increase in lactate dehydrogenase (LDH) and alkaline phosphatase (ALP) leakage, significant inhibition of p-aminohippurate (PAH) accumulation, marked depletion of intracellular ATP and reduced glutathione (GSH), and a significant reduction in pyruvate-stimulated gluconeogenesis. Co-incubation of slices with ADP or CPI and atractyloside completely blocked atractyloside-induced increase in LDH leakage, but not ALP leakage. Atractyloside-induced depletion of ATP and reduced gluconeogenesis was prevented by co-incubation with ADP or CPI. Furthermore, co-incubation of slices with STV and atractyloside, but not PRB, completely abolished atractyloside-induced depletion of ATP and decreased gluconeogenesis in the slices. Pre-incubation of slices with either ADP or CPI protected against atractyloside-induced increase in LDH leakage, reduced ATP and decreased gluconeogenesis. PAH uptake in the slices was inhibited by atractyloside and PRB in a time-dependent manner. While ADP and CPI were found to exert complete protection against atractyloside-induced toxicity irrespective of treatment schedule, STV is effective only under certain conditions, and PRB offer no protection at all. The results of this study demonstrate the usefulness of renal cortical slices as toxicology tool for evaluating and screening compounds for their potential protective effects, and are supportive of a role of adeninine nucleotide (ADP) and protease inhibitor (CPI) in protecting against atractyloside-induced cell injury.

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Microfluidic biochip for the perifusion of precision‐cut rat liver slices for metabolism and toxicology studies

Midwoud

Groothuis

Merema

et al. 2009

Biotech & Bioengineering

123

121

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Early detection of kinetic, metabolic, and toxicity (ADME-Tox) profiles for new drug candidates is of crucial importance during drug development. This article describes a novel in vitro system for the incubation of precision-cut liver slices (PCLS) under flow conditions, based on a poly(dimethylsiloxane) (PDMS) device containing 25-microL microchambers for integration of the slices. The microdevice is coupled to a perifusion system, which enables a constant delivery of nutrients and oxygen and a continuous removal of waste products. Both a highly controlled incubation environment and high metabolite detection sensitivity could be achieved using microfluidics. Liver slices were viable for at least 24 h in the microdevice. The compound, 7-ethoxycoumarin (7-EC), was chosen to test metabolism, since its metabolism includes both phase I and phase II metabolism and when tested in the conventional well plate system, correlates well with the in vivo situation (De Kanter et al. 2004. Xenobiotica 34(3): 229-241.). The metabolic rate of 7-EC was found to be 214 +/- 5 pmol/min/mg protein in the microdevice, comparable to well plates, and was constant over time for at least 3 h. This perifusion system better mimics the in vivo situation, and has the potential to significantly contribute to drug metabolism and toxicology studies of novel chemical entities.

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Optimizing preincubation conditions for precision-cut rat kidney and liver tissue slices: effect of culture media and antioxidants

Cited by 18 publications

References 37 publications

Hydrogel embedding of precision‐cut liver slices in a microfluidic device improves drug metabolic activity

Hydrogel embedding of precision‐cut liver slices in a microfluidic device improves drug metabolic activity

Adenine nucleotide and calpain inhibitor I protect against atractyloside-induced toxicity in rat renal cortical slices in vitro

Microfluidic biochip for the perifusion of precision‐cut rat liver slices for metabolism and toxicology studies

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