Culture duration alters the glutathione content and sensitivity to ethacrynic acid of rat hepatocyte monolayer cultures

1986 ◽  
Vol 2 (4) ◽  
pp. 495-505 ◽  
Author(s):  
Michael J. Meredith
Keyword(s):  
Ethacrynic Acid ◽  
Rat Hepatocyte ◽  
Monolayer Cultures ◽  
Culture Duration ◽  
Hepatocyte Monolayer

Cryopreservation of rat hepatocyte monolayer cultures

1996 ◽  
Vol 15 (1) ◽  
pp. 30-37 ◽  
Author(s):  
P. Watts ◽  
MH Grant
Keyword(s):  
Plasma Membranes ◽  
Rat Hepatocytes ◽  
Repair Process ◽  
Significant Loss ◽  
Urea Synthesis ◽  
Albumin Synthesis ◽  
Rat Hepatocyte ◽  
Monolayer Cultures ◽  
Cryopreservation Method ◽  
Hepatocyte Monolayer

1 Most previous attempts to cryopreserve hepatocytes have used suspensions stored at either - 70°C or in liquid nitrogen, and the major problem is that these do not, on subsequent thawing, attach well in culture. This limits their use in studies of drug metabolism and xenobiotic- induced toxicity. In this manuscript we demonstrate successful cryopreservation of rat hepatocytes as mono layers attached to a collagen film. 2 Monolayers can be frozen and thawed without significant loss of cells, and although damage to the internal and plasma membranes is evident immediately post-thaw, a remarkable repair process takes place over 24-48 h post-thaw. Immediately post-thaw only 10% of the cells exclude Trypan Blue, but by 48 h 80 - 90% of the thawed cells are viable, indicating that repair of the plasma membranes has taken place. 3 The cells post-thaw retain aspects of liver-specific function including cytochrome P450 content and albumin synthesis. However, cytosolic proteins are lost through the damaged membranes and, probably because of this, urea synthesis from ammonia is retained at only 25% of pre- freeze values. 4 A cryopreservation method based on adherent hepa tocytes on a collagen substrate overcomes the problems encountered with culture of cryopreserved hepatocyte suspensions, and may provide a practical means of establishing a 'bank' of hepatocytes from several donors and species.

Regulation of Ketogenesis, Gluconeogenesis, and Glycogen Synthesis by Insulin and Proinsulin in Rat Hepatocyte Monolayer Cultures

Diabetes ◽  
1986 ◽  
Vol 35 (11) ◽  
pp. 1286-1293 ◽  
Author(s):  
L. Agius ◽  
M. H. Chowdhury ◽  
S. N. Davis ◽  
K. G. M. M. Alberti
Keyword(s):  
Glycogen Synthesis ◽  
Rat Hepatocyte ◽  
Monolayer Cultures ◽  
Hepatocyte Monolayer

scholarly journals Regulation of ketogenesis, gluconeogenesis and the mitochondrial redox state by dexamethasone in hepatocyte monolayer cultures

1986 ◽  
Vol 239 (3) ◽  
pp. 593-601 ◽  
Author(s):  
L Agius ◽  
M H Chowdhury ◽  
K G M M Alberti
Keyword(s):  
Redox State ◽  
Ketone Bodies ◽  
Dibutyryl Cyclic Amp ◽  
Pyruvate Metabolism ◽  
Rat Hepatocyte ◽  
Monolayer Cultures ◽  
Mitochondrial Redox State ◽  
Hepatocyte Monolayer ◽  
Parenchymal Hepatocytes ◽  
Reducing Equivalents

The effects of the glucocorticoid dexamethasone on fatty acid and pyruvate metabolism were studied in rat hepatocyte cultures. Parenchymal hepatocytes were cultured for 24 h with nanomolar concentrations of dexamethasone in either the absence or the presence of insulin (10 nM) or dibutyryl cyclic AMP (1 microM BcAMP). Dexamethasone (1-100 nM) increased the rate of formation of ketone bodies from 0.5 mM-palmitate in both the absence and the presence of BcAMP, but inhibited ketogenesis in the presence of insulin. Dexamethasone increased the proportion of the palmitate metabolized that was partitioned towards oxidation to ketone bodies, and decreased the cellular [glycerol 3-phosphate]. The latter suggests that the increased partitioning of palmitate to ketone bodies may be associated with decreased esterification to glycerolipid. The Vmax. of carnitine palmitoyltransferase (CPT) and the affinity of CPT for palmitoyl-CoA were not affected by dexamethasone, indicating that the increased ketogenesis was not due to an increase in enzymic capacity for long-chain acylcarnitine formation. Dexamethasone and BcAMP, separately and in combination, increased gluconeogenesis. In the presence of insulin, however, dexamethasone inhibited gluconeogenesis. Changes in gluconeogenesis thus paralleled changes in ketogenesis. Dexamethasone decreased the [3-hydroxybutyrate]/[acetoacetate] ratio, despite increasing the rate of ketogenesis and presumably the mitochondrial production of reducing equivalents. The more oxidized mitochondrial NADH/NAD+ redox couple with dexamethasone is probably due either to an increased rate of electron transport or to increased transfer of mitochondrial reducing equivalents to the cytoplasm.

Rat Hepatocyte Cultures: Conventional Monolayer Cultures and Cocultures with Rat liver Epithelial Cells

2003 ◽  
pp. 311-318 ◽  
Author(s):  
Tamara Vanhaecke ◽  
Karen De Smet ◽  
Sonja Beken ◽  
Marleen Pauwels ◽  
Antoine Vercruysse ◽  
...  
Keyword(s):  
Epithelial Cells ◽  
Rat Liver ◽  
Rat Hepatocyte ◽  
Liver Epithelial Cells ◽  
Hepatocyte Cultures ◽  
Monolayer Cultures ◽  
Rat Liver Epithelial Cells
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