Lipid and glucose metabolism in hepatocyte cell lines and primary mouse hepatocytes: a comprehensive resource for in vitro studies of hepatic metabolism

Abstract

The liver is a critical tissue for maintaining glucose, fatty acid, and cholesterol homeostasis. Primary hepatocytes represent the gold standard for studying the mechanisms controlling hepatic glucose, lipid, and cholesterol metabolism in vitro. However, access to primary hepatocytes can be limiting, and therefore, other immortalized hepatocyte models are commonly used. Here, we describe substrate metabolism of cultured AML12, IHH, and PH5CH8 cells, hepatocellular carcinoma-derived HepG2s, and primary mouse hepatocytes (PMH) to identify which of these cell lines most accurately phenocopy PMH basal and insulin-stimulated metabolism. Insulin-stimulated glucose metabolism in PH5CH8 cells, and to a lesser extent AML12 cells, responded most similarly to PMH. Notably, glucose incorporation in HepG2 cells were 14-fold greater than PMH. The differences in glucose metabolic activity were not explained by differential protein expression of key regulators of these pathways, for example glycogen synthase and glycogen content. In contrast, fatty acid metabolism in IHH cells was the closest to PMHs, yet insulin-responsive fatty acid metabolism in AML12 and HepG2 cells was most similar to PMH. Finally, incorporation of acetate into intracellular-free cholesterol was comparable for all cells to PMH; however, insulin-stimulated glucose conversion into lipids and the incorporation of acetate into intracellular cholesterol esters were strikingly different between PMHs and all tested cell lines. In general, AML12 cells most closely phenocopied PMH in vitro energy metabolism. However, the cell line most representative of PMHs differed depending on the mode of metabolism being investigated, and so careful consideration is needed in model selection.

Application of In Vitro Metabolism Activation in High-Throughput Screening

“…Cryopreserved hepatocytes [64] are also viable sources of this metabolically competent model and are commercially available. Cryopreserved hepatocytes can lead to successful predictions of in vivo hepatic clearance, indicating an excellent correlation to newly prepared fresh cell cultures [65]. Moreover, the interindividual variability seen with fresh human primary hepatocytes can be irradicated in cryopreserved cells if the same lot of hepatocytes are used for every experiment.…”

Resveratrol Metabolites Are Able to Reduce Steatosis in Cultured Hepatocytes

“…HepG2 is a hepatocellular carcinoma cell line with a gene expression profile that highly correlates with primary hepatocellular carcinoma tumors [ 52 ]. By contrast, nontumoral AML12 cells present an energy metabolism closer to primary hepatocytes [ 53 ].…”

Ameliorating effect of pomegranate peel extract supplement against type 1 diabetes-induced hepatic changes in the rat: biochemical, morphological and ultrastructural microscopic studies

“…Hepatocytes are well-known to be involved in the metabolism of different nutrients including carbohydrate (9).…”