Primary-like Human Hepatocytes Genetically Engineered to Obtain Proliferation Competence as a Capable Application for Energy Metabolism Experiments in In Vitro Oncologic Liver Models
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 t...
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MDPI AG
2022-08-01
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| author | Andrea Scheffschick Jonas Babel Sebastian Sperling Julia Nerusch Natalie Herzog Daniel Seehofer Georg Damm |
| author_facet | Andrea Scheffschick Jonas Babel Sebastian Sperling Julia Nerusch Natalie Herzog Daniel Seehofer Georg Damm |
| author_sort | Andrea Scheffschick |
| collection | DOAJ |
| description | 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. |
| first_indexed | 2024-03-09T11:52:35Z |
| format | Article |
| id | doaj.art-84c07c70c4fd40e4aa7ee51f80a809ab |
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| issn | 2079-7737 |
| language | English |
| last_indexed | 2024-03-09T11:52:35Z |
| publishDate | 2022-08-01 |
| publisher | MDPI AG |
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| series | Biology |
| spelling | doaj.art-84c07c70c4fd40e4aa7ee51f80a809ab2023-11-30T23:13:28ZengMDPI AGBiology2079-77372022-08-01118119510.3390/biology11081195Primary-like Human Hepatocytes Genetically Engineered to Obtain Proliferation Competence as a Capable Application for Energy Metabolism Experiments in In Vitro Oncologic Liver ModelsAndrea Scheffschick0Jonas Babel1Sebastian Sperling2Julia Nerusch3Natalie Herzog4Daniel Seehofer5Georg Damm6Department of Hepatobiliary Surgery and Visceral Transplantation, University Hospital, Leipzig University, 04103 Leipzig, GermanyDepartment of Hepatobiliary Surgery and Visceral Transplantation, University Hospital, Leipzig University, 04103 Leipzig, GermanyDepartment of General, Visceral and Transplantation Surgery, Charité University Medicine Berlin, 13353 Berlin, GermanyDepartment of Hepatobiliary Surgery and Visceral Transplantation, University Hospital, Leipzig University, 04103 Leipzig, GermanyFaculty of Science, Brandenburg University of Technology Cottbus-Senftenberg, 01968 Senftenberg, GermanyDepartment of Hepatobiliary Surgery and Visceral Transplantation, University Hospital, Leipzig University, 04103 Leipzig, GermanyDepartment of Hepatobiliary Surgery and Visceral Transplantation, University Hospital, Leipzig University, 04103 Leipzig, GermanyNon-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.https://www.mdpi.com/2079-7737/11/8/1195upcyte hepatocytesprimary human hepatocytesHepG2 cellsHCCenergy metabolism |
| spellingShingle | Andrea Scheffschick Jonas Babel Sebastian Sperling Julia Nerusch Natalie Herzog Daniel Seehofer Georg Damm Primary-like Human Hepatocytes Genetically Engineered to Obtain Proliferation Competence as a Capable Application for Energy Metabolism Experiments in In Vitro Oncologic Liver Models Biology upcyte hepatocytes primary human hepatocytes HepG2 cells HCC energy metabolism |
| title | Primary-like Human Hepatocytes Genetically Engineered to Obtain Proliferation Competence as a Capable Application for Energy Metabolism Experiments in In Vitro Oncologic Liver Models |
| title_full | Primary-like Human Hepatocytes Genetically Engineered to Obtain Proliferation Competence as a Capable Application for Energy Metabolism Experiments in In Vitro Oncologic Liver Models |
| title_fullStr | Primary-like Human Hepatocytes Genetically Engineered to Obtain Proliferation Competence as a Capable Application for Energy Metabolism Experiments in In Vitro Oncologic Liver Models |
| title_full_unstemmed | Primary-like Human Hepatocytes Genetically Engineered to Obtain Proliferation Competence as a Capable Application for Energy Metabolism Experiments in In Vitro Oncologic Liver Models |
| title_short | Primary-like Human Hepatocytes Genetically Engineered to Obtain Proliferation Competence as a Capable Application for Energy Metabolism Experiments in In Vitro Oncologic Liver Models |
| title_sort | primary like human hepatocytes genetically engineered to obtain proliferation competence as a capable application for energy metabolism experiments in in vitro oncologic liver models |
| topic | upcyte hepatocytes primary human hepatocytes HepG2 cells HCC energy metabolism |
| url | https://www.mdpi.com/2079-7737/11/8/1195 |
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