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Long‐term culture of hepatocytes: Effect of hormones on enzyme activities and metabolic capacity
Author(s) -
Dich John,
Vind Constance,
Grunnet Niels
Publication year - 1988
Publication title -
hepatology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 5.488
H-Index - 361
eISSN - 1527-3350
pISSN - 0270-9139
DOI - 10.1002/hep.1840080109
Subject(s) - glucokinase , glucagon , medicine , pyruvate kinase , endocrinology , insulin , hexokinase , hormone , glucocorticoid , biology , lactate dehydrogenase , chemistry , enzyme , glycolysis , metabolism , biochemistry
(i) Hepatocytes isolated from adult rats were cultured for 2 to 3 weeks on collagen in a modified, serum‐free Waymouth medium containing fatty acids and varying concentrations of glucocorticoid, insulin and glucagon. (ii) In the presence of all three hormones, it was possible to maintain the content of DNA, the activity of glucokinase, pyruvate kinase, hexokinase and lactate dehydrogenase at initial levels for 2 to 3 weeks. The activity of glucokinase and pyruvate kinase was affected by the concentration of insulin. (iii) The activity of alcohol dehydrogenase was stable for 3 days and declined to about 25% of the initial level after 2 weeks of culture, irrespective of the presence of hormones. (iv) Maintenance of albumin secretion was dependent on the presence of glucocorticoid, and glucocorticoid and insulin showed an additive or, at some time points, a synergistic effect on its secretion. (v) The content of cytochrome P‐450 could be kept at 65% of the initial level, provided that a relatively high concentration of dexamethasone was present (10 −6 M ). (vi) In the absence of hormones, urea synthesis was 70% of initial levels throughout the experimental period. With insulin and glucocorticoid present, a high concentration of glucagon (10 −8 M ) was required to maintain the synthesis of urea at this level. (vii) It is concluded that hepatocyte cultures as described in the present study may be a useful, well‐defined system for long‐term metabolic, pharmacologic and toxicologic studies.