BioProject

Huh7.5 cells, which are cultured in human serum containing media instead of FBS containing media, undergo growth arrest and become differentiated. More...

Huh7.5 cells, which are cultured in human serum containing media instead of FBS containing media, undergo growth arrest and become differentiated. In this study we further characterized human serum cultured Huh7.5 cells, particularly with regards to morphology, metabolic profile and hepatocyte specific functions. Similar to primary hepatocytes, HS-cultured Huh7.5 cells become columnar, are polarized, and develop bile canalicular structures on the apical side of the cells. Further analysis by microarray showed that significant transcriptional changes occur in 22-32% of the genes. HS cultured Huh7.5 cells undergo extensive metabolic re-programming. We showed a reversal of a Warburg-like metabolic profile that is typical for proliferating cells. In HS cultured cells large lipid droplets can be found in addition to large glycogen stores. Metabolic modeling shows an increased reliance on beta-oxidation, which was confirmed by an increase in ketone body production, the end products of β-oxidation. Additionally we showed increased conversion of glucose to glycogen. Bile secretion is increased, as are the enzymes involved in the degradation of xenobiotics. Overall these data show that only changing the serum in cell culture media can result in the establishment of a cell type that morphologically, metabolically and functionally resemble primary hepatocytes. Overall design: Samples were analyzed for Huh7.5 cells cultured in fetal bovine serum as well as at different time points – 8 days, 15 days and 23 days – after being transferred to human serum. Biological triplicates were used in each case. Less...