Isolation and culture of adult mouse hepatocytes

WC Li, KL Ralphs, D Tosh - Mouse cell culture: Methods and protocols, 2010 - Springer
WC Li, KL Ralphs, D Tosh
Mouse cell culture: Methods and protocols, 2010Springer
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 …
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.
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