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Related Concept Videos

Cell Culture01:21

Cell Culture

Most vertebrate cells grow in vitro attached to a substrate as a monolayer, called adherent cultures. The flasks and plates used to grow cells are chemically treated to facilitate cell attachment. However, a few cell types, such as hematopoietic cells, can grow in a suspension. In contrast to adherent cultures, suspension cultures can grow in non-treated cultureware using magnetic stirrers or spinner flasks to agitate the culture media

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Induction of an Inflammatory Response in Primary Hepatocyte Cultures from Mice
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Induction of an Inflammatory Response in Primary Hepatocyte Cultures from Mice

Published on: March 10, 2017

Liver cell culture techniques.

José V Castell1, María José Gómez-Lechón

  • 1Unit of Experimental Hepatology, University Hospital "La Fe", Valencia, Spain.

Methods in Molecular Biology (Clifton, N.J.)
|December 20, 2008
PubMed
Summary

Human hepatocytes can be cultured from various liver sources using collagenase perfusion. While short-term cultures maintain liver functions for about a week, optimizing media and coatings is key for extended viability.

Area of Science:

  • Hepatology
  • Cell Culture
  • Biotechnology

Background:

  • Human hepatocytes are crucial for studying liver functions and drug metabolism.
  • Primary hepatocyte cultures are vital but face challenges with limited survival and phenotype maintenance.
  • Various liver tissue sources are available for isolating human hepatocytes.

Purpose of the Study:

  • To detail methods for isolating and culturing human hepatocytes.
  • To establish conditions for maintaining hepatocyte function in short-term cultures.
  • To discuss strategies for improving hepatocyte survival and functionality.

Main Methods:

  • Isolation of hepatocytes using two-step collagenase perfusion from diverse human liver tissues.
  • Culturing hepatocytes in monolayer using chemically defined, serum-free media.

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  • Utilizing extracellular matrix components (Matrigel, collagen, fibronectin) for plate coating.

    • Main Results:

    • High yields of viable human hepatocytes expressing typical hepatic functions can be obtained.
    • Short-term monolayer cultures maintain liver-specific functions for approximately one week.
    • Coating culture plates with extracellular matrix components supports hepatocyte function.

    Conclusions:

    • Established protocols enable short-term culture of human hepatocytes with maintained functionality.
    • Optimized culture conditions, including media and coatings, are essential for hepatocyte research.
    • This chapter provides practical guidance for human hepatocyte culture and functional assessment.