|Hepalife Biosystems, Inc.|
Submitted to: Transactions of the American Society of Artificial Internal Organs
Publication Type: Abstract Only
Publication Acceptance Date: February 15, 2008
Publication Date: March 1, 2008
Citation: Willard, R.R., Meekin, J.H., Talbot, N.C., Caperna, T.J. Characterization of PICM-19H porcine liver stem cell line for potential use in a bioartificial liver. Transactions of the American Society of Artificial Internal Organs 54 (2): 9A. Technical Abstract: A hepatocyte cell line is needed as the biological component of a bioartificial liver (BAL). One candidate is the PICM-19 pig liver stem cell line. These cells have many normal hepatocyte functions often lacking in tumor-derived liver cell lines. The study characterized a PICM-19 derivative cell line, PICM-19H. Following incubation with 5µM 3-methylcholanthrene (3-MC), 50µM rifampicin or 2 mM phenobarbital, the activities of cytochrome (P450) isozymes P450 1A, 2 and 3A were found to be inducible in PICM-19H cells. In comparison to primary adult pig hepatocyte (APH) P450 1A and 3A activities, PICM-19H cells had 30% and 43% of those activities while the tumor-derived human HepG2 C3A cell line had 7% and 0%, respectively. Extensive conjugation, 52% and 96%, of P450 1A and 3A metabolites was found, indicating PICM-19H phase II activities similar to APH. PICM-19H secreted serum proteins, including albumin, alpha fetoprotein, transthyretin, transferrin, apolipoprotein A1, and retinol binding protein. Acetaminophen was toxic to PICM-19H cells, with an LC50 of 12.7± 0.8 mM. Bioactivation of aflatoxin B1 following P450 induction was determined in control and 3-MC treated cultures; PICM-19H cells treated with 3-MC had an LC50 of 1.1 µM compared to controls with an LC50 of 40 µM. Urea production in control cultures was 0.131 µmol N/mg protein/h; addition of 2 mM NH3Cl resulted in an increase in urea production and nearly complete removal of NH3 by 48 h. The PICM-19H cell line, therefore, has the critical metabolic detoxification functions necessary for use in a BAL.