Isolation and culture of adult mouse hepatocytes.
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.
Li
Wan-Chun
WC
Department of Biology and Biochemistry, Centre for Regenerative Medicine, University of Bath, BA2 7AY Bath, UK.
Ralphs
Kate L
KL
Tosh
David
D
D000375:Aging; D000818:Animals; D018929:Cell Culture Techniques / Q000295:instrumentation / Q000379:methods*; D002469:Cell Separation / Q000295:instrumentation / Q000379:methods*; D017364:Collagenases / Q000378:metabolism; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D051379:Mice; D010477:Perfusion
D017364:Collagenases