@misc{HerzogHansenMiethbaueretal., author = {Herzog, Natalie and Hansen, Max and Miethbauer, Sebastian and Schmidtke, Kai-Uwe and Anderer, Ursula and Lupp, Amelie and Sperling, Sebastian and Seehofer, Daniel and Damm, Georg and Scheibner, Katrin and K{\"u}pper, Jan-Heiner}, title = {Primary-like human hepatocytes genetically engineered to obtain proliferation competence display hepatic differentiation characteristics in monolayer and organotypical spheroid cultures}, series = {Cell Biology International}, volume = {40}, journal = {Cell Biology International}, number = {3}, issn = {1095-8355}, doi = {10.1002/cbin.10574}, pages = {341 -- 353}, abstract = {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.}, language = {en} } @misc{KadeHerzogSchmidtkeetal., author = {Kade, S. and Herzog, Natalie and Schmidtke, Kai-Uwe and K{\"u}pper, Jan-Heiner}, title = {Chronic ethanol treatment depletes glutathione regeneration capacity in hepatoma cell line HepG2}, series = {Journal of Cellular Biotechnology}, volume = {1}, journal = {Journal of Cellular Biotechnology}, number = {2}, issn = {2352-3697}, doi = {10.3233/JCB-15019}, pages = {183 -- 190}, language = {en} } @misc{HerzogHansenMiethbaueretal., author = {Herzog, Natalie and Hansen, Max and Miethbauer, Sebastian and Schmidtke, Kai-Uwe and Anderer, Ursula and Lupp, Amelie and Sperling, Sebastian and Seehofer, Daniel and Damm, Georg and Scheibner, Katrin and K{\"u}pper, Jan-Heiner}, title = {Primary like human hepatocytes genetically engineered to obtain proliferation competence display liver biotransformation activity in 2D and 3D culture systems}, series = {Cell Biology International}, volume = {40}, journal = {Cell Biology International}, number = {3}, issn = {1095-8355}, pages = {341 -- 353}, language = {en} } @misc{GeorgeRoedigerSchroederetal., author = {George, Sandra and R{\"o}diger, Stefan and Schr{\"o}der, Christian and Knaut, Michael and K{\"u}pper, Jan-Heiner}, title = {Development of multiplex PCR systems for expression profiling of human cardiomyocytes induced to profilerate by lentivirus transduction of upcyte genes}, series = {Journal of Cellular Biotechnology}, volume = {2}, journal = {Journal of Cellular Biotechnology}, number = {1}, issn = {2352-3697}, doi = {10.3233/JCB-15025}, pages = {35 -- 55}, language = {en} } @misc{MeyerMeyerFiccaKuepper, author = {Meyer, Ralph G. and Meyer-Ficca, Mirella and K{\"u}pper, Jan-Heiner}, title = {Adenoviral vectors for modulation of poly(ADP-ribose) polymerase-1 (PARP1) - dependent DNA repair as a predictive tool for chemotherapy}, series = {Journal of Cellular Biotechnology}, volume = {2}, journal = {Journal of Cellular Biotechnology}, number = {1}, issn = {2352-3697}, doi = {10.3233/JCB-15026}, pages = {57 -- 68}, language = {en} } @misc{HerzogKatzenbergerMartinetal., author = {Herzog, Natalie and Katzenberger, Nadine and Martin, Frank and Schmidtke, Kai-Uwe and K{\"u}pper, Jan-Heiner}, title = {Generation of cytochrome P450 3A4-overexpressing HepG2 cell clones for standardization of hepatocellular testosterone 6β-hydroxylation activity}, series = {Journal of Cellular Biotechnology}, volume = {1}, journal = {Journal of Cellular Biotechnology}, number = {1}, issn = {2352-3697}, doi = {10.3233/JCB-15002}, pages = {15 -- 26}, language = {en} } @misc{GeorgeNoackVaneketal., author = {George, Sandra and Noack, M. and Vanek, Monika and Rentzsch, J. and R{\"o}ber, Nadja and Roggenbuck, Dirk and Conrad, Karsten and K{\"u}pper, Jan-Heiner}, title = {Expression of nicotinic acetylcholine receptor subunits in HEp-2 cells for immunodetection of autoantibody specificities in sera fro Myasthenia gravis patients}, series = {Clinical Hemorheology and Microcirculation}, volume = {61}, journal = {Clinical Hemorheology and Microcirculation}, number = {2}, issn = {1875-8622}, doi = {10.3233/CH-151999}, pages = {385 -- 396}, language = {en} } @misc{GeorgePaulickKnuetteretal., author = {George, Sandra and Paulick, Silvia and Kn{\"u}tter, Ilka and R{\"o}ber, Nadja and Hiemann, Rico and Roggenbuck, Dirk and Conrad, Karsten and K{\"u}pper, Jan-Heiner}, title = {Stable Expression of Human Muscle-Specific Kinase in HEp-2 M4 Cells for Automatic Immunofluorescence Diagnostics of Myasthenia Gravis}, series = {PLOS one}, volume = {9}, journal = {PLOS one}, number = {1}, issn = {1932-6203}, doi = {10.1371/journal.pone.0083924}, pages = {e83924}, language = {en} } @misc{GeorgeGeorgiRoggenbucketal., author = {George, Sandra and Georgi, M. and Roggenbuck, Dirk and Conrad, Karsten and K{\"u}pper, Jan-Heiner}, title = {A strategy for cell-based multiplex diagnostics of Myasthenia gravis and autoimmune encephalitis by modifying the subcellular localization of cell membrane autoantigensgravis}, series = {Clinical Hemorheology and Microcirculation}, volume = {58}, journal = {Clinical Hemorheology and Microcirculation}, number = {1}, issn = {1386-0291}, doi = {10.3233/CH-141897}, pages = {211 -- 228}, language = {en} } @incollection{ConradKuepper, author = {Conrad, Karsten and K{\"u}pper, Jan-Heiner}, title = {Tumor-associated autoantibodies}, series = {Autoantibodies}, booktitle = {Autoantibodies}, editor = {Shoenfeld, Yehuda and Meroni, Pier Luigi and Gershwin, M. Eric}, edition = {3rd edition}, publisher = {Elsevier}, address = {Amsterdam [u.a.]}, isbn = {978-0-444-56378-1}, pages = {275 -- 287}, language = {en} } @misc{NowakMartinGeorgeetal., author = {Nowak, Elisabeth and Martin, F. and George, Sandra and R{\"o}ber, Nadja and Conrad, Karsten and K{\"u}pper, Jan-Heiner}, title = {Bacterial production and purification of immunoreactive paraneoplastic neurological syndrome autoantigen Ma2}, series = {Journal of Cellular Biotechnology}, volume = {2}, journal = {Journal of Cellular Biotechnology}, number = {2}, issn = {2352-3697}, doi = {10.3233/JCB-15029}, pages = {85 -- 91}, abstract = {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.}, language = {en} } @book{PaumenKuepper, author = {Paumen, Anja and K{\"u}pper, Jan-Heiner}, title = {It's the planet, stupid! Sieben Perspektiven zum Klimawandel : mit Interviewbeitr{\"a}gen von Hartmut Graßl, Hans-Werner Sinn, Ernst-Ulrich von Weizs{\"a}cker, Meinhard Miegel, Mathias Binswanger, Enoch zu Guttenberg und Wulf Schiefenh{\"o}vel}, publisher = {oekom Verlag}, address = {M{\"u}nchen}, isbn = {978-3-86581-739-6}, pages = {303}, language = {de} } @inproceedings{GeorgeNoackVaneketal., author = {George, Sandra and Noack, Monika and Vanek, Monika and Rentzsch, Juliane and R{\"o}ber, Nadja and Conrad, Karsten and Roggenbuck, Dirk and K{\"u}pper, Jan-Heiner}, title = {Expression of nicotinic acetylcholine receptor subunits in HEp-2 cells for immunodetection of autoantibody specificities in sera from Myasthenia gravis patients}, series = {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}, booktitle = {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}, publisher = {Pabst Science Publishers}, address = {Lengerich}, isbn = {978-3-95853-104-8}, pages = {195 -- 196}, language = {en} } @misc{KammererKuepper, author = {Kammerer, Sarah and K{\"u}pper, Jan-Heiner}, title = {Human Hepatocyte Systems for in vitro Toxicology Analysis.}, series = {Journal of Cellular Biotechnology}, volume = {3}, journal = {Journal of Cellular Biotechnology}, number = {2}, issn = {2352-3697}, doi = {10.3233/JCB-179012}, pages = {85 -- 93}, language = {en} } @misc{HailekaGeorgeSteinbrechtetal., author = {Haileka, Vanessa and George, Sandra and Steinbrecht, Susanne and Jung, Friedrich and Reinehr, R. and K{\"u}pper, Jan-Heiner}, title = {Colon cancer cells cultured under hyperosmotic conditions as in vitro model to investigate dehydration effects on cancer drug susceptibility}, series = {Clinical Hemorheology and Microcirculation}, volume = {73}, journal = {Clinical Hemorheology and Microcirculation}, number = {1}, issn = {1875-8622}, doi = {10.3233/CH-199210}, pages = {169 -- 176}, language = {en} } @misc{SchulteHubbertKuepperThomasetal., author = {Schulte-Hubbert, Ruth and K{\"u}pper, Jan-Heiner and Thomas, Adam D. and Schrenk, Dieter}, title = {Estragole: DNA adduct formation in primary rat hepatocytes and genotoxic potential in HepG2-CYP1A2 cells}, series = {Toxicology}, volume = {444}, journal = {Toxicology}, issn = {0300-483X}, doi = {10.1016/j.tox.2020.152566}, pages = {8}, abstract = {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.}, language = {en} } @misc{RutzGaoKuepperetal., author = {Rutz, Lukas and Gao, Lan and K{\"u}pper, Jan-Heiner and Schrenk, Dieter}, title = {Structure-dependent genotoxic potencies of selected pyrrolizidine alkaloids in metabolically competent HepG2 cells}, series = {Archives of Toxicology}, volume = {94}, journal = {Archives of Toxicology}, number = {12}, issn = {0340-5761}, doi = {10.1007/s00204-020-02895-z}, pages = {4159 -- 4172}, abstract = {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.}, language = {en} } @misc{KuenzelRauschSchaefferetal., author = {K{\"u}nzel, Stephan R. and Rausch, Johanna S. E. and Sch{\"a}ffer, Charlotte and Hoffmann, Maximilian and K{\"u}nzel, Karolina and Klapproth, Erik and Kant, Theresa and Herzog, Natalie and K{\"u}pper, Jan-Heiner and Lorenz, Kristina and Dudek, Svenja and Emig, Ramona and Ravens, Ursula and Rog-Zielinska, Eva A. and Peyronnet, R{\´e}mi and El-Armouche, Ali}, title = {Modeling atrial fibrosis in vitro - Generation and characterization of a novel human atrial fibroblast cell line}, series = {FEBS Open Bio}, volume = {10}, journal = {FEBS Open Bio}, number = {7}, issn = {2211-5463}, doi = {10.1002/2211-5463.12896}, pages = {1210 -- 1218}, abstract = {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.}, language = {en} } @misc{JungKruegerGengeFrankeetal., author = {Jung, Friedrich and Kr{\"u}ger-Genge, Anne and Franke, Ralf-Peter and Hufert, Frank and K{\"u}pper, Jan-Heiner}, title = {COVID-19 and the endothelium}, series = {Clinical Hemorheology and Microcirculation}, volume = {75}, journal = {Clinical Hemorheology and Microcirculation}, number = {1}, issn = {1875-8622}, doi = {10.3233/CH-209007}, pages = {7 -- 11}, abstract = {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.}, language = {en} } @misc{JungKriegerHufertetal., author = {Jung, Friedrich and Krieger, Volker and Hufert, Frank and K{\"u}pper, Jan-Heiner}, title = {Herd immunity or suppression strategy to combat COVID-19}, series = {Clinical Hemorheology and Microcirculation}, volume = {75}, journal = {Clinical Hemorheology and Microcirculation}, number = {1}, issn = {1875-8622}, doi = {10.3233/CH-209006}, pages = {13 -- 17}, abstract = {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).}, language = {en} }