Isolation of neonatal extrahepatic cholangiocytes

Sara Karjoo, Rebecca G. Wells

Research output: Contribution to journalArticlepeer-review

Abstract

The intra and extrahepatic bile ducts of the liver are developmentally distinct, and may be differentially affected by certain diseases. However, differences between intra and extrahepatic cholangiocytes, and between neonatal and adult cells, are not well understood. Methods for the isolation of cholangiocytes from intrahepatic bile ducts are well established1-4. Isolation of extrahepatic ductal cells, especially from the neonate, has not yet been described, although this would be of great benefit in understanding the differences between distinct cholangiocyte populations and in studying diseases such as biliary atresia that appear to target the extrahepatic ducts. Described here is an optimized technique to isolate both neonatal and adult mouse extrahepatic bile duct cells. This technique yields a pure cell population with minimal contamination from mesenchymal cells like fibroblasts. This method is based on the removal of the extrahepatic ducts and gallbladder, followed by meticulous dissection and scraping to remove fat and fibroblast layers. Structures are embedded in thick layers of collagen and cultured for approximately 3 weeks to allow outgrowth of cholangiocytes in monolayers, which can then be trypsinized and re plated for experimental use.

Original languageEnglish (US)
JournalJournal of Visualized Experiments
Issue number88
DOIs
StatePublished - May 6 2014
Externally publishedYes

Keywords

  • Bile Duct Diseases
  • Bile Ducts
  • Bile duct
  • Biliary atresia
  • Cell culture
  • Common Bile Duct
  • Extrahepatic
  • Fibrosis
  • Gallbladder
  • Issue 88
  • Liver
  • Medicine

ASJC Scopus subject areas

  • Neuroscience(all)
  • Chemical Engineering(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Immunology and Microbiology(all)

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