TY - GEN A1 - Scheffschick, Andrea A1 - Babel, Jonas A1 - Sperling, Sebastian A1 - Nerusch, Julia A1 - Herzog, Natalie A1 - Seehofer, Daniel A1 - Damm, Georg T1 - Primary-like Human Hepatocytes Genetically Engineered to Obtain Proliferation Competence as a Capable Application for Energy Metabolism Experiments in In Vitro Oncologic Liver Models T2 - Biology N2 - Non-alcoholic fatty liver disease (NAFLD), characterized by lipid accumulation in the liver, is the most common cause of liver diseases in Western countries. NAFLD is a major risk factor for developing hepatocellular carcinoma (HCC); however, in vitro evaluation of hepatic cancerogenesis fails due to a lack of liver models displaying a proliferation of hepatocytes. Originally designed to overcome primary human hepatocyte (PHH) shortages, upcyte hepatocytes were engineered to obtain continuous proliferation and, therefore, could be a suitable tool for HCC research. We generated upcyte hepatocytes, termed HepaFH3 cells, and compared their metabolic characteristics to HepG2 hepatoma cells and PHHs isolated from resected livers. For displaying NAFLD-related HCCs, we induced steatosis in all liver models. Lipid accumulation, lipotoxicity and energy metabolism were characterized using biochemical assays and Western blot analysis. We showed that proliferating HepaFH3 cells resemble HepG2, both showing a higher glucose uptake rate, lactate levels and metabolic rate compared to PHHs. Confluent HepaFH3 cells displayed some similarities to PHHs, including higher levels of the transaminases AST and ALT compared to proliferating HepaFH3 cells. We recommend proliferating HepaFH3 cells as a pre-malignant cellular model for HCC research, while confluent HepaFH3 cells could serve as PHH surrogates for energy metabolism studies. KW - upcyte hepatocytes KW - primary human hepatocytes KW - HepG2 cells KW - HCC KW - energy metabolism Y1 - 2022 U6 - https://doi.org/10.3390/biology11081195 SN - 2079-7737 VL - 11 IS - 8 ER - TY - GEN A1 - Nerusch, Julia A1 - Schicht, Gerda A1 - Herzog, Natalie A1 - Küpper, Jan-Heiner A1 - Seehofer, Daniel A1 - Damm, Georg T1 - Investigation and distinction of energy metabolism in proliferating hepatocytes and hepatocellular carcinoma cells T2 - Cells N2 - Metabolic rewiring is a hallmark of both hepatic regeneration and malignant transformation, complicating the identification of cancer-specific traits. This study aimed to distinguish the metabolic profiles of proliferating hepatocytes and hepatocellular carcinoma (HCC) cells through integrated analyses of mRNA and protein expression, along with functional characterization. We compared non-malignant Upcyte® hepatocytes (HepaFH3) cultured under proliferative and confluent conditions with primary human hepatocytes, primary human hepatoma cells, and hepatoma cell lines. Proliferating HepaFH3 cells exhibited features of metabolic reprogramming, including elevated glycolysis, increased HIF1A expression, and ketone body accumulation, while maintaining low c-MYC expression and reduced BDH1 levels, distinguishing them from malignant models. In contrast, HCC cells showed upregulation of HK2, c-MYC, and BDH1, reflecting a shift toward aggressive glycolytic and ketolytic metabolism. Functional assays supported the transcript and protein expression data, demonstrating increased glucose uptake, elevated lactate secretion, and reduced glycogen storage in both proliferating and malignant cells. These findings reveal that cancer-like metabolic changes also occur during hepatic regeneration, limiting the diagnostic utility of individual metabolic markers. HepaFH3 cells thus provide a physiologically relevant in vitro model to study regeneration-associated metabolic adaptation and may offer insights that contribute to distinguishing regenerative from malignant processes. Our findings highlight the potential of integrated metabolic profiling in differentiating proliferation from tumorigenesis. KW - Hepatocellular carcinoma KW - HepaFH3 KW - Metabolic reprogramming KW - Glycolysis KW - Ketone body metabolism KW - Liver regeneration KW - Tumor markers KW - HIF1A KW - c-MYC KW - BDH1 Y1 - 2025 U6 - https://doi.org/10.3390/cells14161254 SN - 2073-4409 VL - 14 IS - 16 SP - 1 EP - 26 PB - MDPI CY - Basel ER -