Search
Author
Title
Vol.
Issue
Year
1st Page

Abstract

 

This article in JAS

  1. Vol. 91 No. 1, p. 66-77
     
    Received: Aug 13, 2012
    Published: December 3, 2014


    3 Corresponding author(s): neil.talbot@ars.usda.gov
 View
 Download
 Share

doi:10.2527/jas.2012-5748

GROWTH AND DEVELOPMENT SYMPOSIUM: Development, characterization, and use of a porcine epiblast-derived liver stem cell line: ARS-PICM-1912

  1. N. C. Talbot 3,
  2. T. J. Caperna and
  3. W. M. Garrett
  1. USDA, ARS, Animal Biosciences and Biotechnology Laboratory, Beltsville Agricultural Research Center, Beltsville, MD 20705

Abstract

Totipotent embryonic stem cell lines have not been established from ungulates; however, we have developed a somatic stem cell line from the in vitro culture of pig epiblast cells. The cell line, ARS-PICM-19, was isolated via colony cloning and was found to spontaneously differentiate into hepatic parenchymal epithelial cell types, namely hepatocytes and bile duct cells. Hepatocytes form as monolayers and bile duct cells as 3-dimensional bile ductules. Transmission electron microscopy revealed that the ductules were composed of radially arranged, monociliated cells with their cilia projecting into the lumen of the ductule whereas hepatocytes were arranged in monolayers with lateral canalicular structures containing numerous microvilli and connected by tight junctions and desmosomes. Extensive Golgi and rough endoplasmic reticulum networks were also present, indicative of active protein synthesis. Analysis of conditioned medium by 2-dimensional electrophoresis and mass spectrometry indicated a spectrum of serum-protein secretion by the hepatocytes. The PICM-19 cell line maintains a range of inducible cytochrome P450 activities and, most notably, is the only nontransformed cell line that synthesizes urea in response to ammonia challenge. The PICM-19 cell line has been used for several biomedical- and agricultural-related purposes, such as the in vitro replication of hepatitis E virus, a zoonotic virus of pigs, and a spaceflight experiment to evaluate somatic stem cell differentiation and liver cell function in microgravity. The cell line was also evaluated as a platform for toxicity testing and has been used in a commercial artificial liver rescue device bioreactor. A PICM-19 subclone, PICM-19H, which only differentiates into hepatocytes, was isolated and methods are currently under development to grow PICM-19 cells without feeder cells. Feeder-cell-independent growth will facilitate the study of mesenchymal–parenchymal interactions that influence the divergent differentiation of the PICM-19 cells, enhance our ability to genetically modify the cells, and provide a better model system to investigate porcine hepatic metabolism.

  Please view the pdf by using the Full Text (PDF) link under 'View' to the left.

Copyright © 2013. American Society of Animal Science