Amino acid supplementation improves cell-specific functions of the rat hepatocytes exposed to human plasma

J. Washizu, C. Chan, F. Berthiaume, R. G. Tompkins, M. Toner, M. L. Yarmush

Research output: Contribution to journalArticle

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Abstract

Maintaining hepatocyte function during plasma exposure is critical for the successful development of hepatocyte-based bioartificial liver assist systems. Past attempts to culture hepatocytes in plasma yielded discouraging results. Using a stable culture model based on sandwiching hepatocytes between two layers of collagen gel, we investigated the effect of hormone and amino acid supplementation during exposure of rat hepatocytes to heparin-treated human plasma for 1 week. Morphology and hepatocyte-specific functions were evaluated for hepatocytes cultured in Dulbecco's Modified Eagle medium (DMEM), nonsupplemented plasma, plasma supplemented with hormones, or with hormones plus amino acids. Amino acids were supplemented at four-fold concentration of Basal Medium Eagle with 4 mM glutamine, whereas hormones included 7.5 μg/mL of hydrocortisone and 50 μU/mL of insulin. Cuboidal structure and bile canaliculi formation were observed throughout the 1-week exposure period for control hepatocytes in DMEM and for hepatocytes cultured in hormone supplemented plasma. Albumin and urea synthesis rates of hepatocytes in hormone plus amino acid supplemented plasma during the last day of plasma exposure were 60.4 ± 13.7 and 75.6 ± 6.5 (μg/day per 1 x 106 cells, mean ± SD), respectively, comparable to cultures in standard culture medium. On the other hand, hepatocytes exposed to nonsupplemented plasma suffered significant morphological and functional damage. The results of this study indicate that hormone plus amino acid supplementation help to restore function in hepatocytes exposed to plasma.

Original languageEnglish (US)
Pages (from-to)497-504
Number of pages8
JournalTissue Engineering
Volume6
Issue number5
DOIs
StatePublished - 2000
Externally publishedYes

Profile

Hepatocytes
Amino Acids
Plasmas
Hormones
Amino acids
Plasma (human)
Rats
Bile Canaliculi
Artificial Liver
Glutamine
Culture Media
Hydrocortisone
Heparin
Urea
Albumins
Collagen
Gels
Insulin
Cortisol
Liver

ASJC Scopus subject areas

  • Biophysics
  • Cell Biology

Cite this

Amino acid supplementation improves cell-specific functions of the rat hepatocytes exposed to human plasma. / Washizu, J.; Chan, C.; Berthiaume, F.; Tompkins, R. G.; Toner, M.; Yarmush, M. L.

In: Tissue Engineering, Vol. 6, No. 5, 2000, p. 497-504.

Research output: Contribution to journalArticle

Washizu, J, Chan, C, Berthiaume, F, Tompkins, RG, Toner, M & Yarmush, ML 2000, 'Amino acid supplementation improves cell-specific functions of the rat hepatocytes exposed to human plasma' Tissue Engineering, vol 6, no. 5, pp. 497-504. DOI: 10.1089/107632700750022143

Washizu, J.; Chan, C.; Berthiaume, F.; Tompkins, R. G.; Toner, M.; Yarmush, M. L. / Amino acid supplementation improves cell-specific functions of the rat hepatocytes exposed to human plasma.

In: Tissue Engineering, Vol. 6, No. 5, 2000, p. 497-504.

Research output: Contribution to journalArticle

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