A possible protective role for free radical scavengers in the culture of rat hepatocytes in serum free medium

1986 ◽  
Vol 3 ◽  
pp. S132
Keyword(s):  
Free Radical ◽  
Rat Hepatocytes ◽  
Protective Role ◽  
Free Radical Scavengers ◽  
Serum Free Medium ◽  
Free Medium ◽  
Radical Scavengers ◽  
Serum Free

Long-term culture of rat hepatocytes on porous membranes in hormonally defined serum-free medium

1993 ◽  
Vol 7 (4) ◽  
pp. 453-459 ◽  
Author(s):  
C. Guery ◽  
J.P. Stepniewski ◽  
B. Vannier ◽  
R. Fournex ◽  
G. Lorenzon
Keyword(s):  
Rat Hepatocytes ◽  
Porous Membranes ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Long Term Culture

scholarly journals Induction of mRNA for phosphoenolpyruvate carboxykinase (GTP) by dexamethasone in cultured rat hepatocytes requires on-going protein synthesis

1987 ◽  
Vol 246 (1) ◽  
pp. 237-240 ◽  
Author(s):  
V L Nebes ◽  
S M Morris
Keyword(s):  
Phosphoenolpyruvate Carboxykinase ◽  
Rat Hepatocytes ◽  
Serum Free Medium ◽  
Free Medium ◽  
Serum Free ◽  
Maximum Induction ◽  
Cultured Rat Hepatocytes ◽  
Time Required ◽  
Necessary And Sufficient ◽  
Stimulation Of

Dexamethasone is necessary and sufficient to induce mRNA for phosphoenolpyruvate carboxykinase (GTP) (PEPCK) by 19-fold in rat hepatocytes cultured in serum-free medium. However, the time required for maximum induction is 16 h. The slow induction suggested that glucocorticoids regulate the expression of an intermediate gene product(s) which is required for glucocorticoid stimulation of PEPCK-gene expression. Consistent with this notion was the finding that cycloheximide completely blocked the response to dexamethasone. In contrast, cycloheximide did not block the response to a cyclic AMP analogue.

Effects of hormones on the maintenance and mitochondrial functions of rat hepatocytes cultured in serum-free medium

1983 ◽  
Vol 61 (7) ◽  
pp. 636-643 ◽  
Author(s):  
E. Jane Wilson ◽  
William C. McMurray
Keyword(s):  
Rat Hepatocytes ◽  
Tyrosine Aminotransferase ◽  
Serum Free Medium ◽  
Free Medium ◽  
Cultured Hepatocytes ◽  
Serum Free ◽  
Glycerophosphate Dehydrogenase ◽  
Mitochondrial Functions ◽  
Gluconeogenic Enzyme ◽  
Glucocorticoid Action

The survival and morphology of rat hepatocytes were examined in primary cell cultures that were maintained in serum-free medium supplemented with different hormones. Insulin and dexamethasone improved survival and maintenance of normal epithelial-shaped cells, although triiodothyronine did not alter cell survival or morphology when added to the medium alone or with other hormones. The level of mitochondrial α-glycerophosphate dehydrogenase and cytochromes a(+ a3), b and c, but not c1, were increased in cultured hepatocytes by triiodothyronine. Although induction of α-glycerophosphate dehydrogenase did not require serum or growth hormone, the triiodothyronine effect was potentiated by insulin plus dexamethasone. This permissive effect of dexamethasone parallels its known glucocorticoid action of increasing the activity of the gluconeogenic enzyme tyrosine aminotransferase in the cultured hepatocytes. Glucagon, which also elevated tyrosine aminotransferase activity, had no effect upon the induction of α-glycerophosphate dehydrogenase by triiodothyronine. Since the culture medium was completely defined and triiodothyronine did not alter survival or morphology of the hepatocytes, the effects upon mitochondrial functions are direct cellular actions of the thyroid hormone.

Receptors for epidermal growth factor (urogastrone) and insulin in primary cultures of rat hepatocytes maintained in serum-free medium

1986 ◽  
Vol 64 (8) ◽  
pp. 803-810 ◽  
Author(s):  
M. O'Connor-McCourt ◽  
M. Soley ◽  
L. J. Hayden ◽  
M. D. Hollenberg
Keyword(s):  
Growth Factor ◽  
Molecular Mass ◽  
Primary Cultures ◽  
Rat Hepatocytes ◽  
Serum Free Medium ◽  
Free Medium ◽  
Cross Link ◽  
Serum Free ◽  
Epidermal Growth

We have analyzed the receptors for epidermal growth factor (urogastrone) (EGF-URO) and insulin in primary cultures of adult rat hepatocytes maintained for up to 3 weeks on human placental cell matrix in serum-free defined medium. Cross-link labeling experiments revealed that the insulin receptor, partially damaged by the collagenase isolation procedure, was rapidly regenerated to yield an intact receptor. In contrast, cross-link labeling of the EGF-URO receptor revealed that, upon prolonged culture, there was a progressive disappearance of the high molecular mass (175 kilodaltons (kDa)) receptor form, and an appearance of low molecular mass receptor species (130 and 105 kDa). After 3 weeks of culture, the low molecular mass receptor forms accounted for all of the labeled EGF-URO receptor present in the cells. Measurements of EGF-URO binding indicated that the number of EGF-URO binding sites per cell (2.0 × 105 ± 0.3 × 105) did not change during the 3 weeks of culture. However, there was a decrease in EGF-URO binding affinity, reflected by an increase in the KD from 0.6 to 3.0 nM. At zero time and after 3 weeks in culture, Scatchard plots of the binding data were linear; at intermediate time points, the plots were curvilinear. Despite the changes in the EGF-URO receptor that occurred, cells were still responsive to EGF-URO in terms of the inhibition of acetate incorporation into lipid. The ED50 for EGF-URO (about 0.2 nM) was the same for short-term cultures (48 h) as for cells maintained in culture for 3 weeks. We conclude that the long-term culture of hepatocytes in serum-free medium yields an altered low molecular form of the EGF-URO receptor that is, nonetheless, functional. The study points to differential changes in receptors for peptide hormones that may occur in long-term hepatocyte cultures and illustrates the feasibility of using such cultures for metabolic studies of the actions of EGF-URO.

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