Skip to main content
Advertisement

Main menu

  • Home
  • Articles
    • Current Issue
    • Fast Forward
    • Latest Articles
    • Archive
  • Information
    • Instructions to Authors
    • Submit a Manuscript
    • FAQs
    • For Subscribers
    • Terms & Conditions of Use
    • Permissions
  • Editorial Board
  • Alerts
    • Alerts
    • RSS Feeds
  • Virtual Issues
  • Feedback
  • Other Publications
    • Drug Metabolism and Disposition
    • Journal of Pharmacology and Experimental Therapeutics
    • Molecular Pharmacology
    • Pharmacological Reviews
    • Pharmacology Research & Perspectives
    • ASPET

User menu

  • My alerts
  • Log in
  • My Cart

Search

  • Advanced search
Drug Metabolism & Disposition
  • Other Publications
    • Drug Metabolism and Disposition
    • Journal of Pharmacology and Experimental Therapeutics
    • Molecular Pharmacology
    • Pharmacological Reviews
    • Pharmacology Research & Perspectives
    • ASPET
  • My alerts
  • Log in
  • My Cart
Drug Metabolism & Disposition

Advanced Search

  • Home
  • Articles
    • Current Issue
    • Fast Forward
    • Latest Articles
    • Archive
  • Information
    • Instructions to Authors
    • Submit a Manuscript
    • FAQs
    • For Subscribers
    • Terms & Conditions of Use
    • Permissions
  • Editorial Board
  • Alerts
    • Alerts
    • RSS Feeds
  • Virtual Issues
  • Feedback
  • Visit dmd on Facebook
  • Follow dmd on Twitter
  • Follow ASPET on LinkedIn
Research ArticleArticle

EXPRESSION AND INDUCTION POTENTIAL OF CYTOCHROMES P450 IN HUMAN CRYOPRESERVED HEPATOCYTES

Dirk Roymans, Pieter Annaert, Jos Van Houdt, Adri Weygers, Jan Noukens, Carlo Sensenhauser, José Silva, Cis Van Looveren, Jan Hendrickx, Geert Mannens and Willem Meuldermans
Drug Metabolism and Disposition July 2005, 33 (7) 1004-1016; DOI: https://doi.org/10.1124/dmd.104.003046
Dirk Roymans
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Pieter Annaert
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jos Van Houdt
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Adri Weygers
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jan Noukens
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Carlo Sensenhauser
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
José Silva
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Cis Van Looveren
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Jan Hendrickx
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Geert Mannens
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
Willem Meuldermans
  • Find this author on Google Scholar
  • Find this author on PubMed
  • Search for this author on this site
  • Article
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF
Loading

Abstract

Fresh human hepatocytes are still considered as the “gold standard” to screen in vitro for cytochrome P450 (P450) induction. However, sparse availability of good quality human liver tissue for research purposes and the demand for standardized cell populations, together with the need for proper storage of the cells not immediately required, have resulted in the development of cryopreservation techniques that provide adequate viability and plateability of hepatocytes after thawing. This study aimed at validating cryopreserved human hepatocytes as a model to investigate P450 induction. Cryopreserved cells from four different donors were plated and cultured for 48 h, followed by incubation in the presence of typical P450 inducers. During the experiments, quality of the cultured cells was monitored both physiologically and morphologically. Concomitantly, the activity of CYP1A2, 2B6, 2C9, 2E1, and 3A4 was measured together with their mRNA and protein expression. Determination of CYP1A2, 2B6, 2C9, 2E1, and 3A4 activity in control versus prototypical inducer-treated hepatocytes revealed a maximal significant mean 11.6-, 2.8-, 1.9-, 1.5-, and 9.0-fold induction over their basal expression, respectively. Protein expression analysis of these P450s confirmed these results. Moreover, a mean 44.9-, 3.5-, 3.2-, and 13.8-fold induction of CYP1A2, 2B6, 2C9, and 3A4 mRNA was observed. Our data demonstrate that cryopreserved human hepatocytes are a valuable tool to study the induction of CYP1A2, 2B6, 2C9, 2E1, and 3A4.

Footnotes

  • Article, publication date, and citation information can be found at http://dmd.aspetjournals.org.

  • doi:10.1124/dmd.104.003046.

  • ABBREVIATIONS: P450, cytochrome P450; NCE, new chemical entity; DMSO, dimethyl sulfoxide; RT, room temperature; BSA, bovine serum albumin; HRP, horseradish peroxidase; HBSS, Hanks' balanced salt solution; PhRMA, Pharmaceutical Research & Manufacturers of America; QRT-PCR, quantitative reverse-transcriptase polymerase chain reaction; PAGE, polyacrylamide gel electrophoresis; MOPS, 3-(N-morpholino)propanesulfonic acid; Bicine, N,N-bis(2-hydroxyethyl)glycine; TBST, Tris-buffered saline/Tween 20.

    • Received November 16, 2004.
    • Accepted March 29, 2005.
  • The American Society for Pharmacology and Experimental Therapeutics
View Full Text

 

DMD articles become freely available 12 months after publication, and remain freely available for 5 years. 

Non-open access articles that fall outside this five year window are available only to institutional subscribers and current ASPET members, or through the article purchase feature at the bottom of the page. 

 

  • Click here for information on institutional subscriptions.
  • Click here for information on individual ASPET membership.

 

Log in using your username and password

Forgot your user name or password?

Purchase access

You may purchase access to this article. This will require you to create an account if you don't already have one.
PreviousNext
Back to top

In this issue

Drug Metabolism and Disposition: 33 (7)
Drug Metabolism and Disposition
Vol. 33, Issue 7
1 Jul 2005
  • Table of Contents
  • About the Cover
  • Index by author
Download PDF
Article Alerts
Sign In to Email Alerts with your Email Address
Email Article

Thank you for sharing this Drug Metabolism & Disposition article.

NOTE: We request your email address only to inform the recipient that it was you who recommended this article, and that it is not junk mail. We do not retain these email addresses.

Enter multiple addresses on separate lines or separate them with commas.
EXPRESSION AND INDUCTION POTENTIAL OF CYTOCHROMES P450 IN HUMAN CRYOPRESERVED HEPATOCYTES
(Your Name) has forwarded a page to you from Drug Metabolism & Disposition
(Your Name) thought you would be interested in this article in Drug Metabolism & Disposition.
CAPTCHA
This question is for testing whether or not you are a human visitor and to prevent automated spam submissions.
Citation Tools
Research ArticleArticle

EXPRESSION AND INDUCTION POTENTIAL OF CYTOCHROMES P450 IN HUMAN CRYOPRESERVED HEPATOCYTES

Dirk Roymans, Pieter Annaert, Jos Van Houdt, Adri Weygers, Jan Noukens, Carlo Sensenhauser, José Silva, Cis Van Looveren, Jan Hendrickx, Geert Mannens and Willem Meuldermans
Drug Metabolism and Disposition July 1, 2005, 33 (7) 1004-1016; DOI: https://doi.org/10.1124/dmd.104.003046

Citation Manager Formats

  • BibTeX
  • Bookends
  • EasyBib
  • EndNote (tagged)
  • EndNote 8 (xml)
  • Medlars
  • Mendeley
  • Papers
  • RefWorks Tagged
  • Ref Manager
  • RIS
  • Zotero
Share
Research ArticleArticle

EXPRESSION AND INDUCTION POTENTIAL OF CYTOCHROMES P450 IN HUMAN CRYOPRESERVED HEPATOCYTES

Dirk Roymans, Pieter Annaert, Jos Van Houdt, Adri Weygers, Jan Noukens, Carlo Sensenhauser, José Silva, Cis Van Looveren, Jan Hendrickx, Geert Mannens and Willem Meuldermans
Drug Metabolism and Disposition July 1, 2005, 33 (7) 1004-1016; DOI: https://doi.org/10.1124/dmd.104.003046
del.icio.us logo Digg logo Reddit logo Twitter logo CiteULike logo Facebook logo Google logo Mendeley logo
  • Tweet Widget
  • Facebook Like
  • Google Plus One

Jump to section

  • Article
    • Abstract
    • Materials and Methods
    • Results
    • Discussion
    • Footnotes
    • References
  • Figures & Data
  • Info & Metrics
  • eLetters
  • PDF

Related Articles

Cited By...

More in this TOC Section

  • BSEP Function in Suspension Hepatocytes
  • Candesartan glucuronide serves as a CYP2C8 inhibitor
  • Role of AADAC on eslicarbazepine acetate hydrolysis
Show more Articles

Similar Articles

  • Home
  • Alerts
Facebook   Twitter   LinkedIn   RSS

Navigate

  • Current Issue
  • Fast Forward by date
  • Fast Forward by section
  • Latest Articles
  • Archive
  • Search for Articles
  • Feedback
  • ASPET

More Information

  • About DMD
  • Editorial Board
  • Instructions to Authors
  • Submit a Manuscript
  • Customized Alerts
  • RSS Feeds
  • Subscriptions
  • Permissions
  • Terms & Conditions of Use

ASPET's Other Journals

  • Journal of Pharmacology and Experimental Therapeutics
  • Molecular Pharmacology
  • Pharmacological Reviews
  • Pharmacology Research & Perspectives
ISSN 1521-009X (Online)

Copyright © 2021 by the American Society for Pharmacology and Experimental Therapeutics