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Extended liver-specific functions of porcine hepatocyte spheroids entrapped in collagen gel

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Abstract

The potential use of porcine hepatocytes in a bioartificial liver device requires large quantities of viable and highly active cells. To facilitate the scaling up of the system, liver specific activities of hepatocytes should be maximized. One way of enhancing the specific activities is to cultivate hepatocytes as multicellular spheroids. Freshly isolated porcine hepatocytes form spheroids when cultivated in suspended cultures. These spheroids exhibit higher activities for a number of liver specific functions compared to hepatocytes cultivated as monolayers. However, these activities decreased in a few days in culture. Entrappment of spheroids in collagen gel sustained their metabolic activities at a stable level over 21 days. Production of albumin and urea by spheroid hepatocytes entrapped in collagen gels were 2 to 3 times higher than those by freshly isolated single cells. P-450 activity was demonstrated by metabolism of lidocaine to its main metabolite, monoethylglycinexylidide. Phase II drug metabolism was demonstrated by glucuronidation of 4-methylumbelliferone. This work shows that porcine hepatocyte spheroids entrapped in collagen maintain differentiated functions for an extended time period. Such hepatocyte spheroid entrappment system may facilitate the development of a bioartificial liver support device.

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Authors and Affiliations

  1. Department of Chemical Engineering and Materials Science, University of Minnesota, 55455-0132, Minneapolis, Minnesota

    Arye Lazar & Wei-Shou Hu

  2. Department of Pharmacy Practice, University of Minnesota, 55455-0132, Minneapolis, Minnesota

    Henry J. Mann

  3. Department of Medicinal Chemistry, University of Minnesota, 55455-0132, Minneapolis, Minnesota

    Rory P. Remmel

  4. Department of Surgery, University of Minnesota, 55455-0132, Minneapolis, Minnesota

    Russell A. Shatford & Frank B. Cerra

  5. Department of Biotechnology, Israel Institute for Biological Research, 70450, Ness-Ziona, Israel

    Arye Lazar

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Lazar, A., Mann, H.J., Remmel, R.P. et al. Extended liver-specific functions of porcine hepatocyte spheroids entrapped in collagen gel. In Vitro Cell Dev Biol - Animal 31, 340–346 (1995). https://doi.org/10.1007/BF02634282

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  • DOI: https://doi.org/10.1007/BF02634282

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