Evaluation of the effect of culture matrices on induction of CYP3A isoforms in cultured porcine hepatocytes

Vinayak P. Hosagrahara, Linda K. Hansen, Gregory J Beilman, Rory P Remmel

Research output: Contribution to journalArticle

11 Citations (Scopus)

Abstract

Several bioartificial liver devices have been developed as temporary therapy for patients suffering from fulminant hepatic failure. Some of these devices contain porcine hepatocytes entrapped in collagen matrices. In order to improve the function of these BAL devices, there exists a need to optimize metabolic function of cultured hepatocytes. The goal of these investigations was to evaluate the effect of altering culture conditions on rifampin- mediated induction of CYP3A isoforms in cultured porcine hepatocytes. Midazolam metabolism was compared in porcine hepatocytes cultured in a monolayer configuration on collagen gels, in a sandwich configuration between collagen gels and a Matrigel overlay, and in spheroidal cultures. The effect of culture conditions was evaluated, by measuring CYP3A-mediated metabolism of midazolam and by immunoblotting to detect CYP3A proteins, in control cultures and in rifampin-treated cultures. Results obtained by normalizing the metabolism rate data to cell numbers (based on DNA content) present at the end of the culture experiment, showed that there was no difference between the different culture conditions tested. Our results suggest that culturing porcine hepatocytes as spheroids or in a sandwich configuration between collagen and Matrigel, offers no advantage in terms of CYP3A-mediated metabolic function on a per cell basis compared to culturing on collagen gels. (C) 2000 Elsevier Science Ireland Ltd.

Original languageEnglish (US)
Pages (from-to)91-106
Number of pages16
JournalChemico-Biological Interactions
Volume127
Issue number1
DOIs
StatePublished - Jun 15 2000

Fingerprint

Cytochrome P-450 CYP3A
Hepatocytes
Protein Isoforms
Swine
Collagen
Metabolism
Gels
Midazolam
Rifampin
Equipment and Supplies
Artificial Liver
Dimercaprol
Acute Liver Failure
Immunoblotting
Liver
Monolayers
Cell Count
DNA
Proteins
Experiments

Keywords

  • CYP3A
  • Culture
  • Hepatocytes
  • Induction
  • Matrigel
  • Midazolam
  • Monolayer
  • Porcine
  • Sand wich
  • Spheroids
  • Spinners

Cite this

Evaluation of the effect of culture matrices on induction of CYP3A isoforms in cultured porcine hepatocytes. / Hosagrahara, Vinayak P.; Hansen, Linda K.; Beilman, Gregory J; Remmel, Rory P.

In: Chemico-Biological Interactions, Vol. 127, No. 1, 15.06.2000, p. 91-106.

Research output: Contribution to journalArticle

@article{123a4f813ac04211840d0f33fa0c63ff,
title = "Evaluation of the effect of culture matrices on induction of CYP3A isoforms in cultured porcine hepatocytes",
abstract = "Several bioartificial liver devices have been developed as temporary therapy for patients suffering from fulminant hepatic failure. Some of these devices contain porcine hepatocytes entrapped in collagen matrices. In order to improve the function of these BAL devices, there exists a need to optimize metabolic function of cultured hepatocytes. The goal of these investigations was to evaluate the effect of altering culture conditions on rifampin- mediated induction of CYP3A isoforms in cultured porcine hepatocytes. Midazolam metabolism was compared in porcine hepatocytes cultured in a monolayer configuration on collagen gels, in a sandwich configuration between collagen gels and a Matrigel overlay, and in spheroidal cultures. The effect of culture conditions was evaluated, by measuring CYP3A-mediated metabolism of midazolam and by immunoblotting to detect CYP3A proteins, in control cultures and in rifampin-treated cultures. Results obtained by normalizing the metabolism rate data to cell numbers (based on DNA content) present at the end of the culture experiment, showed that there was no difference between the different culture conditions tested. Our results suggest that culturing porcine hepatocytes as spheroids or in a sandwich configuration between collagen and Matrigel, offers no advantage in terms of CYP3A-mediated metabolic function on a per cell basis compared to culturing on collagen gels. (C) 2000 Elsevier Science Ireland Ltd.",
keywords = "CYP3A, Culture, Hepatocytes, Induction, Matrigel, Midazolam, Monolayer, Porcine, Sand wich, Spheroids, Spinners",
author = "Hosagrahara, {Vinayak P.} and Hansen, {Linda K.} and Beilman, {Gregory J} and Remmel, {Rory P}",
year = "2000",
month = "6",
day = "15",
doi = "10.1016/S0009-2797(00)00163-0",
language = "English (US)",
volume = "127",
pages = "91--106",
journal = "Chemico-Biological Interactions",
issn = "0009-2797",
publisher = "Elsevier Ireland Ltd",
number = "1",

}

TY - JOUR

T1 - Evaluation of the effect of culture matrices on induction of CYP3A isoforms in cultured porcine hepatocytes

AU - Hosagrahara, Vinayak P.

AU - Hansen, Linda K.

AU - Beilman, Gregory J

AU - Remmel, Rory P

PY - 2000/6/15

Y1 - 2000/6/15

N2 - Several bioartificial liver devices have been developed as temporary therapy for patients suffering from fulminant hepatic failure. Some of these devices contain porcine hepatocytes entrapped in collagen matrices. In order to improve the function of these BAL devices, there exists a need to optimize metabolic function of cultured hepatocytes. The goal of these investigations was to evaluate the effect of altering culture conditions on rifampin- mediated induction of CYP3A isoforms in cultured porcine hepatocytes. Midazolam metabolism was compared in porcine hepatocytes cultured in a monolayer configuration on collagen gels, in a sandwich configuration between collagen gels and a Matrigel overlay, and in spheroidal cultures. The effect of culture conditions was evaluated, by measuring CYP3A-mediated metabolism of midazolam and by immunoblotting to detect CYP3A proteins, in control cultures and in rifampin-treated cultures. Results obtained by normalizing the metabolism rate data to cell numbers (based on DNA content) present at the end of the culture experiment, showed that there was no difference between the different culture conditions tested. Our results suggest that culturing porcine hepatocytes as spheroids or in a sandwich configuration between collagen and Matrigel, offers no advantage in terms of CYP3A-mediated metabolic function on a per cell basis compared to culturing on collagen gels. (C) 2000 Elsevier Science Ireland Ltd.

AB - Several bioartificial liver devices have been developed as temporary therapy for patients suffering from fulminant hepatic failure. Some of these devices contain porcine hepatocytes entrapped in collagen matrices. In order to improve the function of these BAL devices, there exists a need to optimize metabolic function of cultured hepatocytes. The goal of these investigations was to evaluate the effect of altering culture conditions on rifampin- mediated induction of CYP3A isoforms in cultured porcine hepatocytes. Midazolam metabolism was compared in porcine hepatocytes cultured in a monolayer configuration on collagen gels, in a sandwich configuration between collagen gels and a Matrigel overlay, and in spheroidal cultures. The effect of culture conditions was evaluated, by measuring CYP3A-mediated metabolism of midazolam and by immunoblotting to detect CYP3A proteins, in control cultures and in rifampin-treated cultures. Results obtained by normalizing the metabolism rate data to cell numbers (based on DNA content) present at the end of the culture experiment, showed that there was no difference between the different culture conditions tested. Our results suggest that culturing porcine hepatocytes as spheroids or in a sandwich configuration between collagen and Matrigel, offers no advantage in terms of CYP3A-mediated metabolic function on a per cell basis compared to culturing on collagen gels. (C) 2000 Elsevier Science Ireland Ltd.

KW - CYP3A

KW - Culture

KW - Hepatocytes

KW - Induction

KW - Matrigel

KW - Midazolam

KW - Monolayer

KW - Porcine

KW - Sand wich

KW - Spheroids

KW - Spinners

UR - http://www.scopus.com/inward/record.url?scp=0034659111&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=0034659111&partnerID=8YFLogxK

U2 - 10.1016/S0009-2797(00)00163-0

DO - 10.1016/S0009-2797(00)00163-0

M3 - Article

C2 - 10903421

AN - SCOPUS:0034659111

VL - 127

SP - 91

EP - 106

JO - Chemico-Biological Interactions

JF - Chemico-Biological Interactions

SN - 0009-2797

IS - 1

ER -