Comparison of bioenergetic activity of primary porcine hepatocytes cultured in four different media.
Primary hepatocytes have extensively been used in biochemical, pharmacological, and physiological research. Recently, primary porcine hepatocytes have been regarded as the cells of choice for bioartificial liver support systems. The optimum culture medium for hepatocytes to be used in such devices has yet to be defined. In this study we investigated the effectiveness of four culture media in driving energy metabolism of primary porcine hepatocytes. The media selected were William's E medium, medium 1640, medium 199, and hepatocyte medium. Cells (3 x 10(10); viability 87 +/- 6%) were isolated from weanling piglets and seeded on 90-mm plates in the above media supplemented with antibiotics and hormones at a density of 8 x 10(6) viable cells per plate. Using 1H NMR spectroscopy we looked at indices of glycolysis, gluconeogenesis. ketogenesis, and ureagenesis on days 2, 4, and 6 of the experiments (n = 9). We also studied urea and albumin synthesis and total P450 content. The examined metabolic pathways of the hepatocytes were maintained by all media, although there were statistically significant differences between them. All media performed well in glycolysis, ureagenesis, and albumin synthesis. William's E medium and medium 199 outperformed the rest in gluconeogenesis. Medium 199 was best in ketogenesis. Overall, medium 199 was the best at driving energy metabolism from its constituent substrates and we think that it preferentially should be used in the culture of primary porcine hepatocytes.
Dabos
Konstantinos J
KJ
Liver Cell Biology Laboratory, Department of Hepatology, University of Edinburgh, Edinburgh EH16 4SU, Scotland, UK. kostasophia@yahoo.com
Nelson
Leonard J
LJ
Hewage
Chandralal H
CH
Parkinson
John A
JA
Howie
A Forbes
AF
Sadler
Ian H
IH
Hayes
Peter C
PC
Plevris
John N
JN
D000085:Acetates / Q000378:metabolism; D000409:Alanine / Q000378:metabolism; D000418:Albumins / Q000378:metabolism; D000596:Amino Acids / Q000378:metabolism; D000818:Animals; D002470:Cell Survival; D017690:Cell Transplantation / Q000379:methods; D003470:Culture Media / Q000737:chemistry / Q000494:pharmacology*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D004734:Energy Metabolism; D005943:Gluconeogenesis; D005947:Glucose / Q000378:metabolism; D018698:Glutamic Acid / Q000378:metabolism; D006019:Glycolysis; D022781:Hepatocytes / Q000166:cytology* / Q000378:metabolism; D007651:Keto Acids / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D009682:Magnetic Resonance Spectroscopy; D013386:Succinates / Q000378:metabolism; D013552:Swine; D013997:Time Factors; D014508:Urea / Q000378:metabolism
D000085:Acetates; D000418:Albumins; D000596:Amino Acids; D003470:Culture Media; D007651:Keto Acids; D013386:Succinates; D018698:Glutamic Acid; D014508:Urea; D003577:Cytochrome P-450 Enzyme System; D007770:L-Lactate Dehydrogenase; D005947:Glucose; D000409:Alanine