TY - GEN A1 - Herzog, Natalie A1 - Hansen, Max A1 - Miethbauer, Sebastian A1 - Schmidtke, Kai-Uwe A1 - Anderer, Ursula A1 - Lupp, Amelie A1 - Sperling, Sebastian A1 - Seehofer, Daniel A1 - Damm, Georg A1 - Scheibner, Katrin A1 - Küpper, Jan-Heiner T1 - Primary-like human hepatocytes genetically engineered to obtain proliferation competence display hepatic differentiation characteristics in monolayer and organotypical spheroid cultures T2 - Cell Biology International N2 - Primary human hepatocytes are in great demand during drug development and in hepatology. However, both scarcity of tissue supply and donor variability of primary cells create a need for the development of alternative hepatocyte systems. By using a lentivirus vector system to transfer coding sequences of Upcyte® proliferation genes, we generated non-transformed stable hepatocyte cultures from human liver tissue samples. Here, we show data on newly generated proliferation-competent HepaFH3 cells investigated as conventional two-dimensional monolayer and as organotypical three-dimensional (3D) spheroid culture. In monolayer culture, HepaFH3 cells show typical cobblestone-like hepatocyte morphology and anchorage-dependent growth for at least 20 passages. Immunofluorescence staining revealed that characteristic hepatocyte marker proteins cytokeratin 8, human serum albumin, and cytochrome P450 (CYP) 3A4 were expressed. Quantitative real-time PCR analyses showed that expression levels of analyzed phase I CYP enzymes were at similar levels compared to those of cultured primary human hepatocytes and considerably higher than in the liver carcinoma cell line HepG2. Additionally, transcripts for phase II liver enzymes and transporter proteins OATP-C, MRP2, Oct1, and BSEP were present in HepaFH3. The cells produced urea and converted model compounds such as testosterone, diclofenac, and 7-OH-coumarin into phases I and II metabolites. Interestingly, phases I and II enzymes were expressed at about the same levels in convenient monolayer cultures and complex 3D spheroids. In conclusion, HepaFH3 cells and related primary-like hepatocyte lines seem to be promising tools for in vitro research of liver functions and as test system in drug development and toxicology analysis. KW - cytochrome P450 enzyme Y1 - 2016 UR - http://onlinelibrary.wiley.com/doi/10.1002/cbin.10574/full U6 - https://doi.org/10.1002/cbin.10574 SN - 1095-8355 VL - 40 IS - 3 SP - 341 EP - 353 ER - TY - GEN A1 - Kade, S. A1 - Herzog, Natalie A1 - Schmidtke, Kai-Uwe A1 - Küpper, Jan-Heiner T1 - Chronic ethanol treatment depletes glutathione regeneration capacity in hepatoma cell line HepG2 T2 - Journal of Cellular Biotechnology Y1 - 2016 U6 - https://doi.org/10.3233/JCB-15019 SN - 2352-3697 SN - 2352-3689 VL - 1 IS - 2 SP - 183 EP - 190 ER - TY - GEN A1 - Herzog, Natalie A1 - Hansen, Max A1 - Miethbauer, Sebastian A1 - Schmidtke, Kai-Uwe A1 - Anderer, Ursula A1 - Lupp, Amelie A1 - Sperling, Sebastian A1 - Seehofer, Daniel A1 - Damm, Georg A1 - Scheibner, Katrin A1 - Küpper, Jan-Heiner T1 - Primary like human hepatocytes genetically engineered to obtain proliferation competence display liver biotransformation activity in 2D and 3D culture systems T2 - Cell Biology International Y1 - 2016 SN - 1095-8355 SN - 1065-6995 VL - 40 IS - 3 SP - 341 EP - 353 ER - TY - GEN A1 - George, Sandra A1 - Rödiger, Stefan A1 - Schröder, Christian A1 - Knaut, Michael A1 - Küpper, Jan-Heiner T1 - Development of multiplex PCR systems for expression profiling of human cardiomyocytes induced to profilerate by lentivirus transduction of upcyte genes T2 - Journal of Cellular Biotechnology Y1 - 2016 U6 - https://doi.org/10.3233/JCB-15025 SN - 2352-3697 SN - 2352-3689 VL - 2 IS - 1 SP - 35 EP - 55 ER - TY - GEN A1 - Meyer, Ralph G. A1 - Meyer-Ficca, Mirella A1 - Küpper, Jan-Heiner T1 - Adenoviral vectors for modulation of poly(ADP-ribose) polymerase-1 (PARP1) - dependent DNA repair as a predictive tool for chemotherapy T2 - Journal of Cellular Biotechnology Y1 - 2016 U6 - https://doi.org/10.3233/JCB-15026 SN - 2352-3697 SN - 2352-3689 VL - 2 IS - 1 SP - 57 EP - 68 ER - TY - GEN A1 - Herzog, Natalie A1 - Katzenberger, Nadine A1 - Martin, Frank A1 - Schmidtke, Kai-Uwe A1 - Küpper, Jan-Heiner T1 - Generation of cytochrome P450 3A4-overexpressing HepG2 cell clones for standardization of hepatocellular testosterone 6β-hydroxylation activity T2 - Journal of Cellular Biotechnology Y1 - 2015 U6 - https://doi.org/10.3233/JCB-15002 SN - 2352-3697 SN - 2352-3689 VL - 1 IS - 1 SP - 15 EP - 26 ER - TY - GEN A1 - George, Sandra A1 - Noack, M. A1 - Vanek, Monika A1 - Rentzsch, J. A1 - Röber, Nadja A1 - Roggenbuck, Dirk A1 - Conrad, Karsten A1 - Küpper, Jan-Heiner T1 - Expression of nicotinic acetylcholine receptor subunits in HEp-2 cells for immunodetection of autoantibody specificities in sera fro Myasthenia gravis patients T2 - Clinical Hemorheology and Microcirculation Y1 - 2015 U6 - https://doi.org/10.3233/CH-151999 SN - 1875-8622 SN - 1386-0291 VL - 61 IS - 2 SP - 385 EP - 396 ER - TY - GEN A1 - George, Sandra A1 - Paulick, Silvia A1 - Knütter, Ilka A1 - Röber, Nadja A1 - Hiemann, Rico A1 - Roggenbuck, Dirk A1 - Conrad, Karsten A1 - Küpper, Jan-Heiner T1 - Stable Expression of Human Muscle-Specific Kinase in HEp-2 M4 Cells for Automatic Immunofluorescence Diagnostics of Myasthenia Gravis T2 - PLOS one Y1 - 2014 U6 - https://doi.org/10.1371/journal.pone.0083924 SN - 1932-6203 VL - 9 IS - 1 SP - e83924 ER - TY - GEN A1 - George, Sandra A1 - Georgi, M. A1 - Roggenbuck, Dirk A1 - Conrad, Karsten A1 - Küpper, Jan-Heiner T1 - A strategy for cell-based multiplex diagnostics of Myasthenia gravis and autoimmune encephalitis by modifying the subcellular localization of cell membrane autoantigensgravis T2 - Clinical Hemorheology and Microcirculation Y1 - 2014 U6 - https://doi.org/10.3233/CH-141897 SN - 1386-0291 SN - 1875-8622 VL - 58 IS - 1 SP - 211 EP - 228 ER - TY - CHAP A1 - Conrad, Karsten A1 - Küpper, Jan-Heiner ED - Shoenfeld, Yehuda ED - Meroni, Pier Luigi ED - Gershwin, M. Eric T1 - Tumor-associated autoantibodies T2 - Autoantibodies Y1 - 2014 SN - 978-0-444-56378-1 SN - 978-0-444-59377-1 SP - 275 EP - 287 PB - Elsevier CY - Amsterdam [u.a.] ET - 3rd edition ER -