Comparison of Bioenergetic Activity of Primary Porcine Hepatocytes Cultured in Four Different Media

Abstract

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 × 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 × 10 6 viable cells per plate. Using 1 H 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.

Transcription Factors and Medium Suitable for Initiating the Differentiation of Human‐Induced Pluripotent Stem Cells to the Hepatocyte Lineage

“…Drug metabolism is preserved in rat primary hepatocytes cultured in WE [Wu et al, ; Takeba et al, ]. Gluconeogenesis and urea cycle—hepatocyte‐specific functions—are also maintained in WE [Farghali et al, ; Dabos et al, ]. These findings clearly indicate that WE is suitable for hepatocyte culture without loss of hepatocyte‐specific functions.…”

Improved Survival and Initiation of Differentiation of Human Induced Pluripotent Stem Cells to Hepatocyte-Like Cells upon Culture in William’s E Medium followed by Hepatocyte Differentiation Inducer Treatment

“…Rat primary hepatocytes are reportedly capable of maintaining drug metabolism in WE [ 31 , 32 ]. Gluconeogenesis and urea synthesis are also preserved in hepatocytes cultured in WE [ 33 , 34 ]. These studies demonstrate that WE is the most suitable medium for the functional maintenance of cultured hepatocytes, and their results are consistent with those of the present study.…”

Toxicity and intracellular accumulation of bile acids in sandwich-cultured rat hepatocytes: Role of glycine conjugates

“…In the present work, compromised urea formation capacity was chosen as indicator to evaluate the concentration-dependency of in vitro cytotoxicity of BA -added individually or as a mixture -in SCRH. Determination of urea formation by bile acid-exposed hepatocytes enables detecting (sub-lethal) reduction in hepatocyte functionality (Dabos et al 2004;Poll et al 2006). To efficiently measure urea formation in SCRH for our purpose, the well-known urea assay (Goeyens et al 1998) was first miniaturized to a higher throughput 96-well format.…”