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 - TY - GEN A1 - Nowak, Elisabeth A1 - Martin, F. A1 - George, Sandra A1 - Röber, Nadja A1 - Conrad, Karsten A1 - Küpper, Jan-Heiner T1 - Bacterial production and purification of immunoreactive paraneoplastic neurological syndrome autoantigen Ma2 T2 - Journal of Cellular Biotechnology N2 - Paraneoplastic neurological syndromes (PNS) are caused by an immune response against neuronal proteins upon their ectopical expression in tumor cells. Ma2 belongs to the protein family of paraneoplastic Ma antigens (PNMA). Detection of Ma2-specific autoantibodies is relevant for diagnostics of anti-Ma2 PNS and an underlying tumor such as germ testicular cancer, small cell lung cancer or breast cancer. Thus, early tumor treatment should improve the outcome for PNS therapy either. Dot blot immunoassay based on recombinantly expressed and purified autoantigens could offer a sensitive method for identification of paraneoplastic autoantibodies from sera of PNS patients. Here we present purification with IMAC and FPLC of human Ma2 autoantigen upon its recombinant expression in E.coli. Furthermore, we provide evidence that dot blot immunoassays with purified Ma2 autoantigen can be used for detection of Ma2-specific autoantibodies from sera of PNS patients. KW - Autoimmune disease KW - autoantibodies KW - dot blot immunoassay KW - FPLC KW - IMAC KW - Ma2 KW - PNMA2 KW - PNS KW - T7 expression system Y1 - 2017 UR - https://content.iospress.com/articles/journal-of-cellular-biotechnology/jcb15029 U6 - https://doi.org/10.3233/JCB-15029 SN - 2352-3697 VL - 2 IS - 2 SP - 85 EP - 91 ER - TY - BOOK A1 - Paumen, Anja A1 - Küpper, Jan-Heiner T1 - It’s the planet, stupid! Sieben Perspektiven zum Klimawandel : mit Interviewbeiträgen von Hartmut Graßl, Hans-Werner Sinn, Ernst-Ulrich von Weizsäcker, Meinhard Miegel, Mathias Binswanger, Enoch zu Guttenberg und Wulf Schiefenhövel Y1 - 2015 SN - 978-3-86581-739-6 SN - 3-86581-739-4 PB - oekom Verlag CY - München ER - TY - PAT A1 - Küpper, Jan-Heiner T1 - Vorrichtung und Verfahren zur Sequestrierung von atmosphärischen Kohlendioxid Y1 - 2017 N1 - EU-Patentanmeldung ER - TY - CHAP A1 - George, Sandra A1 - Noack, Monika A1 - Vanek, Monika A1 - Rentzsch, Juliane A1 - Röber, Nadja A1 - Conrad, Karsten A1 - Roggenbuck, Dirk A1 - Küpper, Jan-Heiner T1 - Expression of nicotinic acetylcholine receptor subunits in HEp-2 cells for immunodetection of autoantibody specificities in sera from Myasthenia gravis patients T2 - From autoantibody research to standardized diagnostic assays in the management of human diseases, report on the 12th Dresden Symposium on Autoantibodies, September 23-26, 2015 Y1 - 2015 SN - 978-3-95853-104-8 SP - 195 EP - 196 PB - Pabst Science Publishers CY - Lengerich ER - TY - GEN A1 - Krüger-Genge, Anne A1 - Steinbrecht, Susanne A1 - Küpper, Jan-Heiner A1 - Lendlein, Andreas A1 - Jung, Friedrich T1 - Evidence for cytostatic effect of cyclophosphamide on human vein endothelial cells in cancer therapy: preliminary in vitro results. T2 - Clinical Hemorheology and Microcirculation Y1 - 2018 U6 - https://doi.org/10.3233/CH-189125 SN - 1875-8622 SN - 1386-0291 VL - 69 IS - 1-2 SP - 267 EP - 276 ER - TY - GEN A1 - Kammerer, Sarah A1 - Küpper, Jan-Heiner T1 - Human Hepatocyte Systems for in vitro Toxicology Analysis. T2 - Journal of Cellular Biotechnology Y1 - 2018 U6 - https://doi.org/10.3233/JCB-179012 SN - 2352-3697 SN - 2352-3689 VL - 3 IS - 2 SP - 85 EP - 93 ER - TY - GEN A1 - Kammerer, Sarah A1 - Küpper, Jan-Heiner T1 - Optimized protocol for induction of cytochrome P450 enzymes 1A2 and 3A4 in human primary-like hepatocyte cell strain HepaFH3 to study in vitro toxicology T2 - Clinical Hemorheology and Microcirculation Y1 - 2018 U6 - https://doi.org/10.3233/CH-189321 SN - 1386-0291 SN - 1875-8622 VL - 70 IS - 4 SP - 563 EP - 571 ER - TY - GEN A1 - Steinbrecht, Susanne A1 - König, Rosalie A1 - Schmidtke, Kai-Uwe A1 - Herzog, Natalie A1 - Scheibner, Katrin A1 - Krüger-Genge, Anne A1 - Jung, Friedrich A1 - Kammerer, Sarah A1 - Küpper, Jan-Heiner T1 - Metabolic activity testing can underestimate acute drug cytotoxicity as revealed by HepG2 cell clones overexpressing cytochrome P450 2C19 and 3A4 T2 - Toxicology Y1 - 2018 U6 - https://doi.org/10.1016/j.tox.2018.11.008 SN - 0300-483X VL - 412 SP - 37 EP - 47 ER - TY - GEN A1 - Krüger-Genge, Anne A1 - Braune, Steffen A1 - Walter, Maria A1 - Kratz, Karl A1 - Küpper, Jan-Heiner A1 - Krengel, M. A1 - Lendlein, Andreas A1 - Jung, Friedrich T1 - Influence of surface treatments of poly(n-butyl acrylate) networks on fibroblasts adhesion, morphology and viability T2 - Clinical Hemorheology and Microcirculation Y1 - 2018 U6 - https://doi.org/10.3233/CH-189130 SN - 1875-8622 SN - 1386-0291 VL - 69 IS - 1-2 SP - 305 EP - 316 ER - TY - GEN A1 - Nowak, Elisabeth A1 - Kammerer, Sarah A1 - Küpper, Jan-Heiner T1 - ATP-based cell viability assay is superior to trypan blue exclusion and XTT assay in measuring cytotoxicity of anticancer drugs Taxol and Imatinib, and proteasome inhibitor MG-132 on human hepatoma cell line HepG2. T2 - Clinical Hemorheology and Microcirculation Y1 - 2018 U6 - https://doi.org/10.3233/CH-189120 SN - 1386-0291 SN - 1875-8622 VL - 69 IS - 1-2 SP - 327 EP - 336 ER -