Oxygen uptake rates and liver-specific functions of hepatocyte and 3T3 fibroblast co-cultures

Authors

  • Cheul H. Cho,

    1. Center for Engineering in Medicine and Department of Surgery, Massachusetts General Hospital, Harvard Medical School, and the Shriners Hospitals for Children, 51 Blossom St, Boston, Massachusetts 02114; telephone: 617-371-4882; fax: 617-371-4950
    Search for more papers by this author
  • Jaesung Park,

    1. Center for Engineering in Medicine and Department of Surgery, Massachusetts General Hospital, Harvard Medical School, and the Shriners Hospitals for Children, 51 Blossom St, Boston, Massachusetts 02114; telephone: 617-371-4882; fax: 617-371-4950
    Search for more papers by this author
  • Deepak Nagrath,

    1. Center for Engineering in Medicine and Department of Surgery, Massachusetts General Hospital, Harvard Medical School, and the Shriners Hospitals for Children, 51 Blossom St, Boston, Massachusetts 02114; telephone: 617-371-4882; fax: 617-371-4950
    Search for more papers by this author
  • Arno W. Tilles,

    1. Center for Engineering in Medicine and Department of Surgery, Massachusetts General Hospital, Harvard Medical School, and the Shriners Hospitals for Children, 51 Blossom St, Boston, Massachusetts 02114; telephone: 617-371-4882; fax: 617-371-4950
    Search for more papers by this author
  • François Berthiaume,

    1. Center for Engineering in Medicine and Department of Surgery, Massachusetts General Hospital, Harvard Medical School, and the Shriners Hospitals for Children, 51 Blossom St, Boston, Massachusetts 02114; telephone: 617-371-4882; fax: 617-371-4950
    Search for more papers by this author
  • Mehmet Toner,

    1. Center for Engineering in Medicine and Department of Surgery, Massachusetts General Hospital, Harvard Medical School, and the Shriners Hospitals for Children, 51 Blossom St, Boston, Massachusetts 02114; telephone: 617-371-4882; fax: 617-371-4950
    Search for more papers by this author
  • Martin L. Yarmush

    Corresponding author
    1. Center for Engineering in Medicine and Department of Surgery, Massachusetts General Hospital, Harvard Medical School, and the Shriners Hospitals for Children, 51 Blossom St, Boston, Massachusetts 02114; telephone: 617-371-4882; fax: 617-371-4950
    • Center for Engineering in Medicine and Department of Surgery, Massachusetts General Hospital, Harvard Medical School, and the Shriners Hospitals for Children, 51 Blossom St, Boston, Massachusetts 02114; telephone: 617-371-4882; fax: 617-371-4950
    Search for more papers by this author

Abstract

Bioartificial liver (BAL) devices have been developed to treat patients undergoing acute liver failure. One of the most important parameters to consider in designing these devices is the oxygen consumption rate of the seeded hepatocytes which are known to have oxygen consumption rates 10 times higher than most other cell types. Hepatocytes in various culture configurations have been tested in BAL devices including those formats that involve co-culture of hepatocytes with other cell types. In this study, we investigated, for the first time, oxygen uptake rates (OUR)s of hepatocytes co-cultured with 3T3-J2 fibroblasts at various hepatocyte to fibroblast seeding ratios. OURs were determined by measuring the rate of oxygen disappearance using a ruthenium-coated optical probe after closing and sealing the culture dish. Albumin and urea production rates were measured to assess hepatocyte function. Lower hepatocyte density co-cultures demonstrated significantly higher OURs (2 to 3.5-fold) and liver- specific functions (1.6-fold for albumin and 4.5-fold for urea production) on a per cell basis than those seeded at higher densities. Increases in OUR correlated well with increased liver-specific functions. OURs (Vm) were modeled by fitting Michaelis–Menten kinetics and the model predictions closely correlated with the experimental data. This study provides useful information for predicting BAL design parameters that will avoid oxygen limitations, as well as maximize metabolic functions. Biotechnol. Bioeng. 2007;97: 188–199. © 2006 Wiley Periodicals, Inc.

Ancillary