Rabbit hepatocytes in primary culture

Preparation, viability and use in studies of propranolol metabolism

Horace E. Walpole, William M. Lee, Thomas Walle, U. Kristina Walle, Michael J. Wilson, John W. Kennedy

Research output: Contribution to journalArticle

13 Citations (Scopus)

Abstract

Isolated hepatocyte cultures have become a frequently used model system for investigating drug metabolism. Although rat and hamster hepatocytes are frequently used for this purpose, metabolism in these species differs in many respects from human metabolism. A species with a metabolism more closely resembling that of humans might be more useful. Our in vivo experiments demonstrated that the metabolism of propranolol in the rabbit is more similar to that in humans than in rats or hamsters. We therefore examined the usefulness of rabbit parenchymal liver cells for studies of propranolol metabolism. A detailed method is presented for their preparation and culture, along with data on their viability, structure and protein synthesizing capability. One- or two-day-old hepatocyte cultures were exposed to 10 10 μmol/L 3H-propranolol from 30 min to 2 hr; metabolites were identified by gas chromatography-mass spectrometry and quantitated by high-performance liquid chromatography. Propran-olol metabolism was linear over 1 hr, with 15% of the substrate metabolized during this time. The cytochrome P-450 pathways, which result in ring oxidation and side-chain oxidation, were expressed in a reproducible fash-ion similar to that found in vivo in humans. The arylhydrocarbon hydroxylase inhibitor a-naphthoflavone (100 μmol/L) inhibited side-chain oxidation of propranolol by 90% without affecting ring oxidation. In contrast, chlorpromazine (100 μmol/L) was shown to inhibit ring oxidation of propranolol by 85% without affecting side chain oxidation. Cimetidine (250 μmol/L) inhibited both pathways by about 50%. These observations suggest that rabbit hepatocyte metabolic pathways more closely resemble human cell metabolism and thus may be a more useful adjunct than rat or hamster hepatocytes in the study of drug-drug interactions when combined with in vivo studies in humans.

Original languageEnglish (US)
Pages (from-to)394-400
Number of pages7
JournalHepatology
Volume11
Issue number3
StatePublished - Mar 1990

Fingerprint

Propranolol
Hepatocytes
Rabbits
Cricetinae
Cimetidine
Chlorpromazine
Mixed Function Oxygenases
Metabolic Networks and Pathways
Drug Interactions
Pharmaceutical Preparations
Gas Chromatography-Mass Spectrometry
Cytochrome P-450 Enzyme System
High Pressure Liquid Chromatography
Ions
Liver
Proteins

ASJC Scopus subject areas

  • Hepatology

Cite this

Walpole, H. E., Lee, W. M., Walle, T., Walle, U. K., Wilson, M. J., & Kennedy, J. W. (1990). Rabbit hepatocytes in primary culture: Preparation, viability and use in studies of propranolol metabolism. Hepatology, 11(3), 394-400.

Rabbit hepatocytes in primary culture : Preparation, viability and use in studies of propranolol metabolism. / Walpole, Horace E.; Lee, William M.; Walle, Thomas; Walle, U. Kristina; Wilson, Michael J.; Kennedy, John W.

In: Hepatology, Vol. 11, No. 3, 03.1990, p. 394-400.

Research output: Contribution to journalArticle

Walpole, HE, Lee, WM, Walle, T, Walle, UK, Wilson, MJ & Kennedy, JW 1990, 'Rabbit hepatocytes in primary culture: Preparation, viability and use in studies of propranolol metabolism', Hepatology, vol. 11, no. 3, pp. 394-400.
Walpole, Horace E. ; Lee, William M. ; Walle, Thomas ; Walle, U. Kristina ; Wilson, Michael J. ; Kennedy, John W. / Rabbit hepatocytes in primary culture : Preparation, viability and use in studies of propranolol metabolism. In: Hepatology. 1990 ; Vol. 11, No. 3. pp. 394-400.
@article{f399a55366e845e8b1eeaeb977d27f7b,
title = "Rabbit hepatocytes in primary culture: Preparation, viability and use in studies of propranolol metabolism",
abstract = "Isolated hepatocyte cultures have become a frequently used model system for investigating drug metabolism. Although rat and hamster hepatocytes are frequently used for this purpose, metabolism in these species differs in many respects from human metabolism. A species with a metabolism more closely resembling that of humans might be more useful. Our in vivo experiments demonstrated that the metabolism of propranolol in the rabbit is more similar to that in humans than in rats or hamsters. We therefore examined the usefulness of rabbit parenchymal liver cells for studies of propranolol metabolism. A detailed method is presented for their preparation and culture, along with data on their viability, structure and protein synthesizing capability. One- or two-day-old hepatocyte cultures were exposed to 10 10 μmol/L 3H-propranolol from 30 min to 2 hr; metabolites were identified by gas chromatography-mass spectrometry and quantitated by high-performance liquid chromatography. Propran-olol metabolism was linear over 1 hr, with 15{\%} of the substrate metabolized during this time. The cytochrome P-450 pathways, which result in ring oxidation and side-chain oxidation, were expressed in a reproducible fash-ion similar to that found in vivo in humans. The arylhydrocarbon hydroxylase inhibitor a-naphthoflavone (100 μmol/L) inhibited side-chain oxidation of propranolol by 90{\%} without affecting ring oxidation. In contrast, chlorpromazine (100 μmol/L) was shown to inhibit ring oxidation of propranolol by 85{\%} without affecting side chain oxidation. Cimetidine (250 μmol/L) inhibited both pathways by about 50{\%}. These observations suggest that rabbit hepatocyte metabolic pathways more closely resemble human cell metabolism and thus may be a more useful adjunct than rat or hamster hepatocytes in the study of drug-drug interactions when combined with in vivo studies in humans.",
author = "Walpole, {Horace E.} and Lee, {William M.} and Thomas Walle and Walle, {U. Kristina} and Wilson, {Michael J.} and Kennedy, {John W.}",
year = "1990",
month = "3",
language = "English (US)",
volume = "11",
pages = "394--400",
journal = "Hepatology",
issn = "0270-9139",
publisher = "John Wiley and Sons Ltd",
number = "3",

}

TY - JOUR

T1 - Rabbit hepatocytes in primary culture

T2 - Preparation, viability and use in studies of propranolol metabolism

AU - Walpole, Horace E.

AU - Lee, William M.

AU - Walle, Thomas

AU - Walle, U. Kristina

AU - Wilson, Michael J.

AU - Kennedy, John W.

PY - 1990/3

Y1 - 1990/3

N2 - Isolated hepatocyte cultures have become a frequently used model system for investigating drug metabolism. Although rat and hamster hepatocytes are frequently used for this purpose, metabolism in these species differs in many respects from human metabolism. A species with a metabolism more closely resembling that of humans might be more useful. Our in vivo experiments demonstrated that the metabolism of propranolol in the rabbit is more similar to that in humans than in rats or hamsters. We therefore examined the usefulness of rabbit parenchymal liver cells for studies of propranolol metabolism. A detailed method is presented for their preparation and culture, along with data on their viability, structure and protein synthesizing capability. One- or two-day-old hepatocyte cultures were exposed to 10 10 μmol/L 3H-propranolol from 30 min to 2 hr; metabolites were identified by gas chromatography-mass spectrometry and quantitated by high-performance liquid chromatography. Propran-olol metabolism was linear over 1 hr, with 15% of the substrate metabolized during this time. The cytochrome P-450 pathways, which result in ring oxidation and side-chain oxidation, were expressed in a reproducible fash-ion similar to that found in vivo in humans. The arylhydrocarbon hydroxylase inhibitor a-naphthoflavone (100 μmol/L) inhibited side-chain oxidation of propranolol by 90% without affecting ring oxidation. In contrast, chlorpromazine (100 μmol/L) was shown to inhibit ring oxidation of propranolol by 85% without affecting side chain oxidation. Cimetidine (250 μmol/L) inhibited both pathways by about 50%. These observations suggest that rabbit hepatocyte metabolic pathways more closely resemble human cell metabolism and thus may be a more useful adjunct than rat or hamster hepatocytes in the study of drug-drug interactions when combined with in vivo studies in humans.

AB - Isolated hepatocyte cultures have become a frequently used model system for investigating drug metabolism. Although rat and hamster hepatocytes are frequently used for this purpose, metabolism in these species differs in many respects from human metabolism. A species with a metabolism more closely resembling that of humans might be more useful. Our in vivo experiments demonstrated that the metabolism of propranolol in the rabbit is more similar to that in humans than in rats or hamsters. We therefore examined the usefulness of rabbit parenchymal liver cells for studies of propranolol metabolism. A detailed method is presented for their preparation and culture, along with data on their viability, structure and protein synthesizing capability. One- or two-day-old hepatocyte cultures were exposed to 10 10 μmol/L 3H-propranolol from 30 min to 2 hr; metabolites were identified by gas chromatography-mass spectrometry and quantitated by high-performance liquid chromatography. Propran-olol metabolism was linear over 1 hr, with 15% of the substrate metabolized during this time. The cytochrome P-450 pathways, which result in ring oxidation and side-chain oxidation, were expressed in a reproducible fash-ion similar to that found in vivo in humans. The arylhydrocarbon hydroxylase inhibitor a-naphthoflavone (100 μmol/L) inhibited side-chain oxidation of propranolol by 90% without affecting ring oxidation. In contrast, chlorpromazine (100 μmol/L) was shown to inhibit ring oxidation of propranolol by 85% without affecting side chain oxidation. Cimetidine (250 μmol/L) inhibited both pathways by about 50%. These observations suggest that rabbit hepatocyte metabolic pathways more closely resemble human cell metabolism and thus may be a more useful adjunct than rat or hamster hepatocytes in the study of drug-drug interactions when combined with in vivo studies in humans.

UR - http://www.scopus.com/inward/record.url?scp=0025361676&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0025361676&partnerID=8YFLogxK

M3 - Article

VL - 11

SP - 394

EP - 400

JO - Hepatology

JF - Hepatology

SN - 0270-9139

IS - 3

ER -