Protocol

Mouse Cell Culture

Volume 633 of the series Methods in Molecular Biology pp 185-196

Date:

Isolation and Culture of Adult Mouse Hepatocytes

  • Wan-Chun LiAffiliated withDepartment of Biology and Biochemistry, Centre for Regenerative Medicine, University of Bath
  • , Kate L. RalphsAffiliated withDepartment of Biology and Biochemistry, Centre for Regenerative Medicine, University of Bath
  • , David ToshAffiliated withDepartment of Biology and Biochemistry, Centre for Regenerative Medicine, University of Bath

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Abstract

The liver performs a multitude of functions including the regulation of carbohydrate, fat, and protein metabolism, the detoxification of endo- and xenobiotics, and the synthesis and secretion of plasma proteins and bile. Isolated hepatocytes constitute a useful technique for studying liver function in an in vitro setting. Here we describe a method for the isolation of hepatocytes from adult mouse liver. The principle of the method is the two-step collagenase perfusion technique which involves sequential perfusion of the liver with ethylenediaminetetraacetic acid and collagenase. Following isolation, the cells can either be used for short-term studies or, alternatively, maintained in culture for prolonged periods to study long-term changes in gene expression. The protocol for mouse hepatocyte isolation may be applied to both normal and transgenic mice.

Key words

Hepatocytes primary culture transgenic collagenase