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Isolation and Culturing Primary Chaolangiocytes from Mouse Liver.

Bio-protocol (2021-11-12)
Ramesh Kudira, Bal Krishan Sharma, Mary Mullen, Sujit K Mohanty, Bryan Donnelly, Gregory M Tiao, Alexander Miethke
ABSTRACT

Cholangiocytes are epithelial cells lining the intrahepatic and extrahepatic bile ducts. Cholangiocytes perform key physiological functions in the liver. Bile synthesized by hepatocytes is secreted into bile canaliculi, further stored in the gallbladder, and finally discharged into the duodenum. Due to liver injury, biliary epithelial proliferate in response to endogenous or exogenous signals leading to cholangiopathies, inflammation, fibrosis, and cholangiocarcinoma. Cholangiocytes exhibit anatomical and functional heterogeneity, and understanding such diversified functions will potentially help in finding effective therapies for various cholestatic liver diseases. To perform such functional studies, effective cholangiocyte isolation and culture procedures are needed. This protocol will aid in easy isolation and expansion of cholangiocytes from the liver.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
Hyaluronidase from bovine testes, Type I-S, lyophilized powder, 400-1000 units/mg solid
Sigma-Aldrich
Forskolin, For use in molecular biology applications
Sigma-Aldrich
Monoclonal Anti-β-Actin antibody produced in mouse, clone AC-15, ascites fluid