Optimization of rat hepatocyte culture in citrated human plasma

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

Research output: Contribution to journalArticle

  • 27 Citations

Abstract

Background. Maintenance of liver-specific functions in hepatocyte cultures during plasma exposure is critical for the clinical application of bioartificial liver assist systems. Sodium citrate is a common anticoagulant but has been shown to be cytotoxic to hepatocytes. We have tested the effect of various supplements on the viability and function of adult primary rat hepatocytes exposed to citrated plasma. Materials and methods. Freshly isolated rat hepatocytes were cultured in the collagen gel sandwich configuration in culture medium for 6 days followed by exposure to citrated human plasma with various supplements for 1 week. Controls were left in culture medium throughout. Viability and synthetic functions were evaluated. Results. Hepatocytes exposed to unsupplemented citrated plasma lost significant viability and function within the first 2 days. Cells cultured in plasma supplemented with a fivefold concentrate of standard hepatocyte culture medium maintained urea (1.2-2.1 μmol/day/106 cells) and albumin (51-62 μg/day/106 cells) synthesis rates equal to or higher than those of controls. Among the various components of the concentrated medium supplement, calcium chloride (1.8 mM), magnesium sulfate (0.8 mM), amino acids (four-fold Basal Medium Eagle amino acids including 4 mM glutamine), and glucagon (14 ng/ml) were found to be essential in maintaining urea synthesis. Maintenance of a high albumin synthesis rate also required the addition of hydrocortisone (7.5 μg/ml) and insulin (0.5 U/ml). Conclusions. Appropriate metabolic and hormonal supplementation of citrated human plasma prevents its cytotoxic effects and may be used in conjunction with in vivo use of bioartificial liver assist systems. (C) 2000 Academic Press.

Original languageEnglish (US)
Pages (from-to)237-246
Number of pages10
JournalJournal of Surgical Research
Volume93
Issue number2
DOIs
StatePublished - 2000
Externally publishedYes

Profile

Hepatocytes
Culture Media
Artificial Liver
Urea
Albumins
Amino Acids
Magnesium Sulfate
Calcium Chloride
Glucagon
Glutamine
Citric Acid
Anticoagulants
Hydrocortisone
Cultured Cells
Collagen
Gels
Sodium
Insulin
Liver

Keywords

  • Amino acids
  • Anticoagulant
  • Bioartificial liver assist system
  • Hepatocyte culture
  • Hormonal supplementation
  • Plasma exposure

ASJC Scopus subject areas

  • Surgery

Cite this

Washizu, J., Berthiaume, F., Chan, C., Tompkins, R. G., Toner, M., & Yarmush, M. L. (2000). Optimization of rat hepatocyte culture in citrated human plasma. Journal of Surgical Research, 93(2), 237-246. DOI: 10.1006/jsre.2000.5986

Optimization of rat hepatocyte culture in citrated human plasma. / Washizu, J.; Berthiaume, F.; Chan, C.; Tompkins, R. G.; Toner, M.; Yarmush, M. L.

In: Journal of Surgical Research, Vol. 93, No. 2, 2000, p. 237-246.

Research output: Contribution to journalArticle

Washizu, J, Berthiaume, F, Chan, C, Tompkins, RG, Toner, M & Yarmush, ML 2000, 'Optimization of rat hepatocyte culture in citrated human plasma' Journal of Surgical Research, vol 93, no. 2, pp. 237-246. DOI: 10.1006/jsre.2000.5986
Washizu J, Berthiaume F, Chan C, Tompkins RG, Toner M, Yarmush ML. Optimization of rat hepatocyte culture in citrated human plasma. Journal of Surgical Research. 2000;93(2):237-246. Available from, DOI: 10.1006/jsre.2000.5986

Washizu, J.; Berthiaume, F.; Chan, C.; Tompkins, R. G.; Toner, M.; Yarmush, M. L. / Optimization of rat hepatocyte culture in citrated human plasma.

In: Journal of Surgical Research, Vol. 93, No. 2, 2000, p. 237-246.

Research output: Contribution to journalArticle

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