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 - 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 - 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 - Haileka, Vanessa A1 - George, Sandra A1 - Steinbrecht, Susanne A1 - Jung, Friedrich A1 - Reinehr, R. A1 - Küpper, Jan-Heiner T1 - Colon cancer cells cultured under hyperosmotic conditions as in vitro model to investigate dehydration effects on cancer drug susceptibility T2 - Clinical Hemorheology and Microcirculation Y1 - 2019 U6 - https://doi.org/10.3233/CH-199210 SN - 1875-8622 VL - 73 IS - 1 SP - 169 EP - 176 ER - TY - GEN A1 - Schulte-Hubbert, Ruth A1 - Küpper, Jan-Heiner A1 - Thomas, Adam D. A1 - Schrenk, Dieter T1 - Estragole: DNA adduct formation in primary rat hepatocytes and genotoxic potential in HepG2-CYP1A2 cells T2 - Toxicology N2 - Estragole is a natural constituent in herbs and spices and in products thereof such as essential oils or herbal teas. After cytochrome P450-catalyzed hydroxylation and subsequent sulfation, estragole acts as a genotoxic hepatocarcinogen forming DNA adducts in rodent liver. Because of the genotoxic mode of action and the widespread occurrence in food and phytomedicines a refined risk assessment for estragole is needed. We analyzed the time- and concentration-dependent levels of the DNA adducts N2-(isoestragole-3‘-yl)-2‘-desoxyguanosine (E3′N2dG) and N6-(isoestragole-3‘-yl)-desoxyadenosine (E3′N6dA), reported to be the major adducts formed in rat liver, in rat hepatocytes (pRH) in primary culture after incubation with estragole. DNA adduct levels were measured via UHPLC-ESI-MS/MS using stable isotope dilution analysis. Both adducts were formed in pRH and could already be quantified after an incubation time of 1 h (E3′N6dA at 10 μM, E3′N2dG at 1μM estragole). E3′N2dG, the main adduct at all incubation times and concentrations, could be detected at estragole concentrations < 0.1 μM after 24 h and < 0.5 μM after 48 h. Adduct levels were highest after 6 h and showed a downward trend at later time-points, possibly due to DNA repair and/or apoptosis. While the concentration-response characteristics of adduct formation were apparently linear over the whole concentration range, strong indication for marked hypo-linearity was obtained when the modeling was based on concentrations < 1 μM only. In the micronucleus assay no mutagenic potential of estragole was found in HepG2 cells whereas in HepG2-CYP1A2 cells 1 μM estragole led to a 3.2 fold and 300 μM to a 7.1 fold increase in micronuclei counts. Our findings suggest the existence of a ‘practical threshold’ dose for DNA adduct formation as an initiating key event of the carcinogenicity of estragole indicating that the default assumption of concentration-response-linearity is questionable, at least for the two major adducts studied here. KW - Estragole KW - Carcinogenicity KW - DNA adducts KW - Genotoxicity KW - Hepatoma KW - Liver cells Y1 - 2020 UR - https://www.sciencedirect.com/science/article/pii/S0300483X20302055 U6 - https://doi.org/10.1016/j.tox.2020.152566 SN - 0300-483X VL - 444 ER - TY - GEN A1 - Rutz, Lukas A1 - Gao, Lan A1 - Küpper, Jan-Heiner A1 - Schrenk, Dieter T1 - Structure-dependent genotoxic potencies of selected pyrrolizidine alkaloids in metabolically competent HepG2 cells T2 - Archives of Toxicology N2 - 1,2-unsaturated pyrrolizidine alkaloids (PAs) are natural plant constituents comprising more than 600 different structures. A major source of human exposure is thought to be cross-contamination of food, feed and phytomedicines with PA plants. In humans, laboratory and farm animals, certain PAs exert pronounced liver toxicity and can induce malignant liver tumors in rodents. Here, we investigated the cytotoxicity and genotoxicity of eleven PAs belonging to different structural classes. Although all PAs were negative in the fluctuation Ames test in Salmonella, they were cytotoxic and induced micronuclei in human HepG2 hepatoblastoma cells over-expressing human cytochrome P450 3A4. Lasiocarpine and cyclic diesters except monocrotaline were the most potent congeners both in cytotoxicity and micronucleus assays with concentrations below 3 μM inducing a doubling in micronuclei counts. Other open di-esters and all monoesters exhibited weaker or much weaker geno- and cytotoxicity. The findings were in agreement with recently suggested interim Relative Potency (iREP) factors with the exceptions of europine and monocrotaline. A more detailed micronuclei analysis at low concentrations of lasiocarpine, retrorsine or senecionine indicated that pronounced hypolinearity of the concentration–response curves was evident for retrorsine and senecionine but not for lasiocarpine. Our findings show that the genotoxic and cytotoxic potencies of PAs in a human hepatic cell line vary in a structure-dependent manner. Both the low potency of monoesters and the shape of prototype concentration–response relationships warrant a substance- and structure-specific approach in the risk assessment of PAs. KW - Genotoxicity KW - Liver cells KW - Micronuclei KW - Mutagenicity KW - Pyrrolizidine alkaloids KW - Relative potencies Y1 - 2020 UR - https://link.springer.com/article/10.1007/s00204-020-02895-z U6 - https://doi.org/10.1007/s00204-020-02895-z SN - 0340-5761 SN - 1432-0738 VL - 94 IS - 12 SP - 4159 EP - 4172 ER - TY - GEN A1 - Künzel, Stephan R. A1 - Rausch, Johanna S. E. A1 - Schäffer, Charlotte A1 - Hoffmann, Maximilian A1 - Künzel, Karolina A1 - Klapproth, Erik A1 - Kant, Theresa A1 - Herzog, Natalie A1 - Küpper, Jan-Heiner A1 - Lorenz, Kristina A1 - Dudek, Svenja A1 - Emig, Ramona A1 - Ravens, Ursula A1 - Rog‐Zielinska, Eva A. A1 - Peyronnet, Rémi A1 - El‐Armouche, Ali T1 - Modeling atrial fibrosis in vitro - Generation and characterization of a novel human atrial fibroblast cell line T2 - FEBS Open Bio N2 - Atrial fibrillation (AF) is regularly accompanied by cardiac fibrosis and concomitant heart failure. Due to the heterogeneous nature and complexity of fibrosis, the knowledge about the underlying mechanisms is limited, which prevents effective pharmacotherapy. A deeper understanding of cardiac fibroblasts is essential to meet this need. We previously described phenotypic and functional differences between atrial fibroblasts from patients in sinus rhythm and with AF. Herein, we established and characterized a novel human atrial fibroblast line, which displays typical fibroblast morphology and function comparable to primary cells but with improved proliferation capacity and low spontaneous myofibroblast differentiation. These traits make our model suitable for the study of fibrosis mechanisms and for drug screening aimed at developing effective antifibrotic pharmacotherapy. KW - cardiovascular disease KW - cell culture KW - cell lines KW - fibroblasts KW - fibrosis KW - heart Y1 - 2020 U6 - https://doi.org/10.1002/2211-5463.12896 SN - 2211-5463 VL - 10 IS - 7 SP - 1210 EP - 1218 ER - TY - GEN A1 - Jung, Friedrich A1 - Krüger-Genge, Anne A1 - Franke, Ralf-Peter A1 - Hufert, Frank A1 - Küpper, Jan-Heiner T1 - COVID-19 and the endothelium T2 - Clinical Hemorheology and Microcirculation N2 - There is growing evidence that COVID-19 not only affects the lungs but beyond that the endothelial system. Recent studies showed that this can lead to microcirculatory impairments and in consequence to functional disorders of all inner organs. The combination of endothelial dysfunction with a generalized inflammatory state and complement elements may together contribute to the overall pro-coagulative state described in COVID-19 patients leading to venular as well as to arteriolar occlusions. KW - COVID-19 KW - endocytosis KW - endothelial cell dysfunction KW - endotheliitis Y1 - 2020 UR - https://content.iospress.com/articles/clinical-hemorheology-and-microcirculation/ch209007 U6 - https://doi.org/10.3233/CH-209007 SN - 1875-8622 SN - 1386-0291 VL - 75 IS - 1 SP - 7 EP - 11 ER - TY - GEN A1 - Jung, Friedrich A1 - Krieger, Volker A1 - Hufert, Frank A1 - Küpper, Jan-Heiner T1 - Herd immunity or suppression strategy to combat COVID-19 T2 - Clinical Hemorheology and Microcirculation N2 - Some months ago, severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) broke out in Wuhan, China, and spread rapidly around the world. Some states, such as the Netherlands, Germany, Great Britain, Sweden and the USA initially focused on keeping the restrictions for economy and society as low as possible. The responsible authorities were of the opinion - and still are e.g. in Sweden - that it is sufficient enough to protect particularly vulnerable persons such as the elderly or people with pre-existing conditions. The idea behind this is that as soon as 60 to 70 percent of the population is infected with a pathogen, a so-called “herd immunity” has developed. However, the increasing numbers of deaths and modelling studies showed the expected overload of the hospitals. Therefore, most countries decided for a temporary lockdown with the exception of Sweden. Based on the number of the total population, three times more people died from COVID-19 in Sweden (2679 deaths per 10 million inhabitants) compared to Germany (6848 deaths per 80 million inhabitants). The comparison Sweden versus Taiwan is even worse because 1072 times more people died in Sweden based on the number of the population (6 deaths per 24 million inhabitants). KW - COVID-19 KW - strategy KW - Sweden KW - Taiwan KW - Germany Y1 - 2020 UR - https://content.iospress.com/articles/clinical-hemorheology-and-microcirculation/ch209006 U6 - https://doi.org/10.3233/CH-209006 SN - 1875-8622 SN - 1386-0291 VL - 75 IS - 1 SP - 13 EP - 17 ER -