Journal
JOVE-JOURNAL OF VISUALIZED EXPERIMENTS
Volume -, Issue 157, Pages -Publisher
JOURNAL OF VISUALIZED EXPERIMENTS
DOI: 10.3791/60992
Keywords
Medicine; Issue 157; liver; slices; vibratome; ex vivo; hepatocytes; tissue; culture; stellate cells
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Funding
- National Health and Medical Research Council (NHMRC) of Australia [APP1048740, APP1142394, APP1160323]
- NHMRC of Australia [APP1061332]
- TALENTO program of Madrid, Spain [T1-BIO-1854]
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Understanding the mechanisms of liver injury, hepatic fibrosis, and cirrhosis that underlie chronic liver diseases (i.e., viral hepatitis, non-alcoholic fatty liver disease, metabolic liver disease, and liver cancer) requires experimental manipulation of animal models and in vitro cell cultures. Both techniques have limitations, such as the requirement of large numbers of animals for in vivo manipulation. However, in vitro cell cultures do not reproduce the structure and function of the multicellular hepatic environment. The use of precision-cut liver slices is a technique in which uniform slices of viable mouse liver are maintained in laboratory tissue culture for experimental manipulation. This technique occupies an experimental niche that exists between animal studies and in vitro cell culture methods. The presented protocol describes a straightforward and reliable method to isolate and culture precision-cut liver slices from mice. As an application of this technique, ex vivo liver slices are treated with bile acids to simulate cholestatic liver injury and ultimately assess the mechanisms of hepatic fibrogenesis.
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