Isolation and culture of adult mouse hepatocytes

Methods Mol Biol. 2010;633:185-96. doi: 10.1007/978-1-59745-019-5_13.

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.

MeSH terms

  • Aging*
  • Animals
  • Cell Culture Techniques / instrumentation
  • Cell Culture Techniques / methods*
  • Cell Separation / instrumentation
  • Cell Separation / methods*
  • Collagenases / metabolism
  • Hepatocytes / cytology*
  • Hepatocytes / metabolism
  • Mice
  • Perfusion

Substances

  • Collagenases