{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,16]],"date-time":"2026-10-16T18:28:07Z","timestamp":1792175287631,"version":"4.4.0"},"reference-count":69,"publisher":"MDPI AG","issue":"10","license":[{"start":{"date-parts":[[2018,10,12]],"date-time":"2018-10-12T00:00:00Z","timestamp":1539302400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Stiftung zur F\u00f6rderung der Erforschung von Ersatz- und Erg\u00e4nzungsmethoden zur Einschr\u00e4nkung von Tierversuchen (SET)","award":["1"],"award-info":[{"award-number":["1"]}]},{"DOI":"10.13039\/501100008751","name":"Bundesinstitut f\u00fcr Risikobewertung","doi-asserted-by":"publisher","award":["1328-568"],"award-info":[{"award-number":["1328-568"]}],"id":[{"id":"10.13039\/501100008751","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["SFB-TR84 B06"],"award-info":[{"award-number":["SFB-TR84 B06"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001659","name":"Deutsche Forschungsgemeinschaft","doi-asserted-by":"publisher","award":["SFB-TR84 Z01"],"award-info":[{"award-number":["SFB-TR84 Z01"]}],"id":[{"id":"10.13039\/501100001659","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Bioprinting is a novel technology that may help to overcome limitations associated with two-dimensional (2D) cell cultures and animal experiments, as it allows the production of three-dimensional (3D) tissue models composed of human cells. The present study describes the optimization of a bioink composed of alginate, gelatin and human extracellular matrix (hECM) to print human HepaRG liver cells with a pneumatic extrusion printer. The resulting tissue model was tested for its suitability for the study of transduction by an adeno-associated virus (AAV) vector and infection with human adenovirus 5 (hAdV5). We found supplementation of the basic alginate\/gelatin bioink with 0.5 and 1 mg\/mL hECM provides desirable properties for the printing process, the stability of the printed constructs, and the viability and metabolic functions of the printed HepaRG cells. The tissue models were efficiently transduced by AAV vectors of serotype 6, which successfully silenced an endogenous target (cyclophilin B) by means of RNA interference. Furthermore, the printed 3D model supported efficient adenoviral replication making it suitable to study virus biology and develop new antiviral compounds. We consider the approach described here paradigmatic for the development of 3D tissue models for studies including viral vectors and infectious viruses.<\/jats:p>","DOI":"10.3390\/ijms19103129","type":"journal-article","created":{"date-parts":[[2018,10,12]],"date-time":"2018-10-12T02:58:04Z","timestamp":1539313084000},"page":"3129","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":133,"title":["Generation of a 3D Liver Model Comprising Human Extracellular Matrix in an Alginate\/Gelatin-Based Bioink by Extrusion Bioprinting for Infection and Transduction Studies"],"prefix":"10.3390","volume":"19","author":[{"given":"Thomas","family":"Hiller","sequence":"first","affiliation":[{"name":"Institute of Biotechnology, Department of Applied Biochemistry, Technische Universit\u00e4t Berlin, 13355 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Johanna","family":"Berg","sequence":"additional","affiliation":[{"name":"Institute of Biotechnology, Department of Applied Biochemistry, Technische Universit\u00e4t Berlin, 13355 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Laura","family":"Elomaa","sequence":"additional","affiliation":[{"name":"Institute of Chemistry and Biochemistry, Department of Organic Chemistry, Freie Universit\u00e4t Berlin, 14195 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Viola","family":"R\u00f6hrs","sequence":"additional","affiliation":[{"name":"Institute of Biotechnology, Department of Applied Biochemistry, Technische Universit\u00e4t Berlin, 13355 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Imran","family":"Ullah","sequence":"additional","affiliation":[{"name":"Berlin-Brandenburger Centrum f\u00fcr Regenerative Therapien, Charit\u00e9 - Universit\u00e4tsmedizin Berlin, 13353 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Katrin","family":"Schaar","sequence":"additional","affiliation":[{"name":"Institute of Biotechnology, Department of Applied Biochemistry, Technische Universit\u00e4t Berlin, 13355 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ann-Christin","family":"Dietrich","sequence":"additional","affiliation":[{"name":"Institute of Biotechnology, Department of Applied Biochemistry, Technische Universit\u00e4t Berlin, 13355 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Munir A.","family":"Al-Zeer","sequence":"additional","affiliation":[{"name":"Institute of Biotechnology, Department of Applied Biochemistry, Technische Universit\u00e4t Berlin, 13355 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas","family":"Kurtz","sequence":"additional","affiliation":[{"name":"Berlin-Brandenburger Centrum f\u00fcr Regenerative Therapien, Charit\u00e9 - Universit\u00e4tsmedizin Berlin, 13353 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Andreas C.","family":"Hocke","sequence":"additional","affiliation":[{"name":"Dept. of Internal Medicine\/Infectious and Respiratory Diseases, Charit\u00e9 \u2212 Universit\u00e4tsmedizin Berlin, 10117 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Stefan","family":"Hippenstiel","sequence":"additional","affiliation":[{"name":"Dept. of Internal Medicine\/Infectious and Respiratory Diseases, Charit\u00e9 \u2212 Universit\u00e4tsmedizin Berlin, 10117 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Henry","family":"Fechner","sequence":"additional","affiliation":[{"name":"Institute of Biotechnology, Department of Applied Biochemistry, Technische Universit\u00e4t Berlin, 13355 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Marie","family":"Weinhart","sequence":"additional","affiliation":[{"name":"Institute of Chemistry and Biochemistry, Department of Organic Chemistry, Freie Universit\u00e4t Berlin, 14195 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-1469-0052","authenticated-orcid":false,"given":"Jens","family":"Kurreck","sequence":"additional","affiliation":[{"name":"Institute of Biotechnology, Department of Applied Biochemistry, Technische Universit\u00e4t Berlin, 13355 Berlin, Germany"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2018,10,12]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1038\/laban0704-37","article-title":"Cell culture and animal models of viral hepatitis. Part I: Hepatitis B","volume":"33","author":"Guha","year":"2004","journal-title":"Lab. Anim. (NY)"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1097\/01.mph.0000153958.95486.6f","article-title":"Fatal adenovirus hepatitis during standard chemotherapy for childhood acute lymphoblastic leukemia","volume":"27","author":"Hough","year":"2005","journal-title":"J. Pediatr. Hematol. Oncol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"1530","DOI":"10.3390\/v20801530","article-title":"Animal Models for Influenza Virus Pathogenesis and Transmission","volume":"2","author":"Bouvier","year":"2010","journal-title":"Viruses"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"300","DOI":"10.1016\/j.omtn.2017.07.002","article-title":"Anti-adenoviral Artificial MicroRNAs Expressed from AAV9 Vectors Inhibit Human Adenovirus Infection in Immunosuppressed Syrian Hamsters","volume":"8","author":"Schaar","year":"2017","journal-title":"Mol. Ther. Nucleic Acids"},{"key":"ref_5","unstructured":"Knipe, D.M., and Howley, P.M. (2007). Adenoviridae: The viruses and Their Replication. Fields Virology, Lippinocott Williams & Wilkins."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"1333","DOI":"10.1038\/sj.bmt.1702716","article-title":"Outcome and clinical course of 100 patients with adenovirus infection following bone marrow transplantation","volume":"26","author":"Baldwin","year":"2000","journal-title":"Bone Marrow Transplant."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"316","DOI":"10.1038\/nrg2971","article-title":"State-of-the-art gene-based therapies: The road ahead","volume":"12","author":"Kay","year":"2011","journal-title":"Nat. Rev. Genet."},{"key":"ref_8","unstructured":"(2018). Voretigene neparvovec-rzyl (Luxturna) for inherited retinal dystrophy. Med. Lett. Drugs Ther., 60, 53\u201355."},{"key":"ref_9","first-page":"1831","article-title":"Endgame: Glybera finally recommended for approval as the first gene therapy drug in the European union","volume":"20","year":"2010","journal-title":"Mol. Ther."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"59","DOI":"10.1016\/j.coph.2015.07.006","article-title":"Adeno-associated virus serotypes for gene therapeutics","volume":"24","author":"Lisowski","year":"2015","journal-title":"Curr. Opin. Pharmacol."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"1115","DOI":"10.1089\/104303402753812511","article-title":"Adeno-associated viral vectors penetrate human solid tumor tissue in vivo more effectively than adenoviral vectors","volume":"13","author":"Enger","year":"2002","journal-title":"Hum. Gene Ther."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"1131","DOI":"10.1002\/jgm.939","article-title":"Adeno-associated virus (AAV) serotypes 2, 4 and 5 display similar transduction profiles and penetrate solid tumor tissue in models of human glioma","volume":"8","author":"Thorsen","year":"2006","journal-title":"J. Gene Med."},{"key":"ref_13","doi-asserted-by":"crossref","unstructured":"Grix, T., Ruppelt, A., Thomas, A., Amler, A.K., Noichl, B.P., Lauster, R., and Kloke, L. (2018). Bioprinting Perfusion-Enabled Liver Equivalents for Advanced Organ-on-a-Chip Applications. Genes (Basel), 9.","DOI":"10.3390\/genes9040176"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1257","DOI":"10.1016\/j.drudis.2016.04.006","article-title":"Application areas of 3D bioprinting","volume":"21","author":"Ozbolat","year":"2016","journal-title":"Drug Discov. Today"},{"key":"ref_15","doi-asserted-by":"crossref","unstructured":"You, F., Eames, B.F., and Chen, X. (2017). Application of Extrusion-Based Hydrogel Bioprinting for Cartilage Tissue Engineering. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18071597"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"83","DOI":"10.1089\/ten.2006.12.83","article-title":"Generation of three-dimensional hepatocyte\/gelatin structures with rapid prototyping system","volume":"12","author":"Wang","year":"2006","journal-title":"Tissue Eng."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1016\/j.cryobiol.2010.10.161","article-title":"Incorporation of DMSO and dextran-40 into a gelatin\/alginate hydrogel for controlled assembled cell cryopreservation","volume":"61","author":"Wang","year":"2010","journal-title":"Cryobiology"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"49","DOI":"10.1016\/j.biomaterials.2013.09.078","article-title":"The 3D printing of gelatin methacrylamide cell-laden tissue-engineered constructs with high cell viability","volume":"35","author":"Billiet","year":"2014","journal-title":"Biomaterials"},{"key":"ref_19","doi-asserted-by":"crossref","unstructured":"Axpe, E., and Oyen, M.L. (2016). Applications of Alginate-Based Bioinks in 3D Bioprinting. Int. J. Mol. Sci., 17.","DOI":"10.3390\/ijms17121976"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"217","DOI":"10.1016\/j.biotechadv.2016.12.006","article-title":"The bioink: A comprehensive review on bioprintable materials","volume":"35","author":"Hospodiuk","year":"2017","journal-title":"Biotechnol. Adv."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"787","DOI":"10.1016\/j.tibtech.2018.03.003","article-title":"Organ-Derived Decellularized Extracellular Matrix: A Game Changer for Bioink Manufacturing?","volume":"36","author":"Choudhury","year":"2018","journal-title":"Trends Biotechnol."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1229","DOI":"10.1021\/acs.biomac.6b01908","article-title":"Development of Liver Decellularized Extracellular Matrix Bioink for Three-Dimensional Cell Printing-Based Liver Tissue Engineering","volume":"18","author":"Lee","year":"2017","journal-title":"Biomacromolecules"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"6188","DOI":"10.1021\/bi00267a025","article-title":"Isolation and characterization of type IV procollagen, laminin, and heparan sulfate proteoglycan from the EHS sarcoma","volume":"21","author":"Kleinman","year":"1982","journal-title":"Biochemistry"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"1886","DOI":"10.1002\/pmic.200900758","article-title":"Matrigel: A complex protein mixture required for optimal growth of cell culture","volume":"10","author":"Hughes","year":"2010","journal-title":"Proteomics"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"378","DOI":"10.1016\/j.semcancer.2005.05.004","article-title":"Matrigel: Basement membrane matrix with biological activity","volume":"15","author":"Kleinman","year":"2005","journal-title":"Semin. Cancer Biol."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"3","DOI":"10.1016\/j.addr.2014.06.005","article-title":"Matrigel: From discovery and ECM mimicry to assays and models for cancer research","volume":"79\u201380","author":"Benton","year":"2014","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_27","first-page":"55596","article-title":"Extracellular Matrix and 3D Printing","volume":"2","author":"Eo","year":"2017","journal-title":"Curr. Trends Biomed. Eng. Biosci."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"905","DOI":"10.1517\/14712598.2012.685714","article-title":"Proliferative human cell sources applied as biocomponent in bioartificial livers: A review","volume":"12","author":"Nibourg","year":"2012","journal-title":"Expert Opin. Biol. Ther."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"75","DOI":"10.1124\/dmd.105.006759","article-title":"Expression of cytochromes P450, conjugating enzymes and nuclear receptors in human hepatoma HepaRG cells","volume":"34","author":"Aninat","year":"2006","journal-title":"Drug Metab. Dispos."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1444","DOI":"10.1124\/dmd.107.020016","article-title":"Evaluation of HepaRG cells as an in vitro model for human drug metabolism studies","volume":"36","author":"Kanebratt","year":"2008","journal-title":"Drug Metab. Dispos."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"261","DOI":"10.1007\/978-1-60761-688-7_13","article-title":"The HepaRG cell line: Biological properties and relevance as a tool for cell biology, drug metabolism, and virology studies","volume":"640","author":"Marion","year":"2010","journal-title":"Methods Mol. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","unstructured":"Rahali, K., Ben Messaoud, G., Kahn, C.J.F., Sanchez-Gonzalez, L., Kaci, M., Cleymand, F., Fleutot, S., Linder, M., Desobry, S., and Arab-Tehrany, E. (2017). Synthesis and Characterization of Nanofunctionalized Gelatin Methacrylate Hydrogels. Int. J. Mol. Sci., 18.","DOI":"10.3390\/ijms18122675"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1080\/00914037.2016.1201830","article-title":"3D printing of porous alginate\/gelatin hydrogel scaffolds and their mechanical property characterization","volume":"66","author":"You","year":"2016","journal-title":"Int. J. Pol. Mater. Pol. Biomater."},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1016\/j.matdes.2018.03.002","article-title":"3D printing of concentrated alginate\/gelatin scaffolds with homogeneous nano apatite coating for bone tissue engineering","volume":"146","author":"Lou","year":"2018","journal-title":"Mater. Des."},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"13877","DOI":"10.1038\/s41598-018-31880-x","article-title":"Optimization of cell-laden bioinks for 3D bioprinting and efficient infection with influenza A virus","volume":"8","author":"Berg","year":"2018","journal-title":"Sci. Rep."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"318","DOI":"10.1128\/CMR.05011-11","article-title":"Infection barriers to successful xenotransplantation focusing on porcine endogenous retroviruses","volume":"25","author":"Denner","year":"2012","journal-title":"Clin. Microbiol. Rev."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1197","DOI":"10.1038\/nmeth.3619","article-title":"Tissue matrix arrays for high-throughput screening and systems analysis of cell function","volume":"12","author":"Beachley","year":"2015","journal-title":"Nat. Methods"},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"1531","DOI":"10.1016\/j.addr.2003.08.002","article-title":"Collagens\u2014Structure, function, and biosynthesis","volume":"55","author":"Gelse","year":"2003","journal-title":"Adv. Drug Deliv. Rev."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"6425","DOI":"10.1016\/j.biomaterials.2010.04.064","article-title":"The differential regulation of cell motile activity through matrix stiffness and porosity in three dimensional collagen matrices","volume":"31","author":"Seemann","year":"2010","journal-title":"Biomaterials"},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"683","DOI":"10.1089\/ten.teb.2014.0086","article-title":"Review of collagen I hydrogels for bioengineered tissue microenvironments: Characterization of mechanics, structure, and transport","volume":"20","author":"Antoine","year":"2014","journal-title":"Tissue Eng. Part. B Rev."},{"key":"ref_41","first-page":"a004978","article-title":"The collagen family","volume":"3","year":"2011","journal-title":"Cold Spring Harb. Perspect. Biol."},{"key":"ref_42","unstructured":"Turkse, K. (2015). Bioprinting in Regenrative Medicine, Springer."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"3707","DOI":"10.1016\/j.biomaterials.2003.10.052","article-title":"Inkjet printing for high-throughput cell patterning","volume":"25","author":"Roth","year":"2004","journal-title":"Biomaterials"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"24474","DOI":"10.1038\/srep24474","article-title":"Bioprinting three-dimensional cell-laden tissue constructs with controllable degradation","volume":"6","author":"Wu","year":"2016","journal-title":"Sci. Rep."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1089\/ten.tec.2013.0335","article-title":"Design and fabrication of human skin by three-dimensional bioprinting","volume":"20","author":"Lee","year":"2014","journal-title":"Tissue Eng. Part. C Methods"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"035004","DOI":"10.1088\/1758-5082\/6\/3\/035004","article-title":"A comparative study on collagen type I and hyaluronic acid dependent cell behavior for osteochondral tissue bioprinting","volume":"6","author":"Park","year":"2014","journal-title":"Biofabrication"},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"8596","DOI":"10.1016\/j.biomaterials.2010.07.072","article-title":"Dense type I collagen matrices that support cellular remodeling and microfabrication for studies of tumor angiogenesis and vasculogenesis in vitro","volume":"31","author":"Cross","year":"2010","journal-title":"Biomaterials"},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"854015","DOI":"10.1155\/2015\/854015","article-title":"Interactions between CYP3A4 and Dietary Polyphenols","volume":"2015","author":"Basheer","year":"2015","journal-title":"Oxid. Med. Cell. Longev."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"495","DOI":"10.15171\/apb.2016.063","article-title":"Overview of Albumin and Its Purification Methods","volume":"6","author":"Raoufinia","year":"2016","journal-title":"Adv. Pharm. Bull."},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"15655","DOI":"10.1073\/pnas.232137699","article-title":"Infection of a human hepatoma cell line by hepatitis B virus","volume":"99","author":"Gripon","year":"2002","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"1147","DOI":"10.1053\/j.gastro.2004.01.002","article-title":"Origin and characterization of a human bipotent liver progenitor cell line","volume":"126","author":"Parent","year":"2004","journal-title":"Gastroenterology"},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"1483","DOI":"10.1016\/j.biocel.2011.06.011","article-title":"The HepaRG cell line is suitable for bioartificial liver application","volume":"43","author":"Hoekstra","year":"2011","journal-title":"Int. J. Biochem. Cell. Biol."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"20140817","DOI":"10.1098\/rsif.2014.0817","article-title":"Natural polymers for the microencapsulation of cells","volume":"11","author":"Gasperini","year":"2014","journal-title":"J. R. Soc. Interface"},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"97","DOI":"10.1093\/toxsci\/kfn109","article-title":"Cellular imaging predictions of clinical drug-induced liver injury","volume":"105","author":"Xu","year":"2008","journal-title":"Toxicol. Sci."},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"407","DOI":"10.1124\/dmd.111.039297","article-title":"Differential modulation of cytochrome P450 activity and the effect of 1-aminobenzotriazole on hepatic transport in sandwich-cultured human hepatocytes","volume":"40","author":"Kimoto","year":"2012","journal-title":"Drug Metab. Dispos."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"1936","DOI":"10.1021\/acs.chemrestox.6b00150","article-title":"Novel 3D Culture Systems for Studies of Human Liver Function and Assessments of the Hepatotoxicity of Drugs and Drug Candidates","volume":"29","author":"Lauschke","year":"2016","journal-title":"Chem. Res. Toxicol."},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"8060","DOI":"10.1039\/C7TB01458A","article-title":"The development of cell-initiated degradable hydrogel based on methacrylated alginate applicable to multiple microfabrication technologies","volume":"5","author":"Sun","year":"2017","journal-title":"J. Mater. Chem. B"},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1093\/toxsci\/kft021","article-title":"3D organotypic cultures of human HepaRG cells: A tool for in vitro toxicity studies","volume":"133","author":"Gunness","year":"2013","journal-title":"Toxicol. Sci."},{"key":"ref_59","doi-asserted-by":"crossref","first-page":"e00208","DOI":"10.1042\/BSR20150034","article-title":"3D spheroid cultures improve the metabolic gene expression profiles of HepaRG cells","volume":"35","author":"Takahashi","year":"2015","journal-title":"Biosci. Rep."},{"key":"ref_60","doi-asserted-by":"crossref","first-page":"2164","DOI":"10.1016\/j.biomaterials.2008.12.084","article-title":"Organ printing: Tissue spheroids as building blocks","volume":"30","author":"Mironov","year":"2009","journal-title":"Biomaterials"},{"key":"ref_61","doi-asserted-by":"crossref","first-page":"8475","DOI":"10.3390\/molecules16108475","article-title":"Pharmacological and biological antiviral therapeutics for cardiac coxsackievirus infections","volume":"16","author":"Fechner","year":"2011","journal-title":"Molecules"},{"key":"ref_62","doi-asserted-by":"crossref","first-page":"251","DOI":"10.1038\/424251b","article-title":"Looking into the safety of AAV vectors","volume":"424","author":"Kay","year":"2003","journal-title":"Nature"},{"key":"ref_63","doi-asserted-by":"crossref","first-page":"287","DOI":"10.1016\/j.jconrel.2016.01.001","article-title":"Adeno-associated virus (AAV) vectors in cancer gene therapy","volume":"240","author":"Schaffer","year":"2016","journal-title":"J. Control. Release"},{"key":"ref_64","doi-asserted-by":"crossref","first-page":"281","DOI":"10.2174\/1566523034578285","article-title":"From virus evolution to vector revolution: Use of naturally occurring serotypes of adeno-associated virus (AAV) as novel vectors for human gene therapy","volume":"3","author":"Grimm","year":"2003","journal-title":"Curr. Gene Ther."},{"key":"ref_65","doi-asserted-by":"crossref","first-page":"473","DOI":"10.1182\/asheducation-2007.1.473","article-title":"RNAi and Gene Therapy: A Mutual Attraction","volume":"2007","author":"Grimm","year":"2007","journal-title":"Hematol. Am. Soc. Hematol. Educ. Program."},{"key":"ref_66","doi-asserted-by":"crossref","first-page":"107","DOI":"10.1182\/blood-2005-12-5115","article-title":"Multiyear therapeutic benefit of AAV serotypes 2, 6, and 8 delivering factor VIII to hemophilia A mice and dogs","volume":"108","author":"Jiang","year":"2006","journal-title":"Blood"},{"key":"ref_67","first-page":"e54633","article-title":"Study of Viral Vectors in a Three-dimensional Liver Model Repopulated with the Human Hepatocellular Carcinoma Cell Line HepG2","volume":"116","author":"Hiller","year":"2016","journal-title":"J. Vis. Exp."},{"key":"ref_68","doi-asserted-by":"crossref","first-page":"134","DOI":"10.1016\/j.jbiotec.2015.08.012","article-title":"Use of a three-dimensional humanized liver model for the study of viral gene vectors","volume":"212","author":"Wagner","year":"2015","journal-title":"J. Biotechnol."},{"key":"ref_69","doi-asserted-by":"crossref","first-page":"355","DOI":"10.1089\/hgtb.2013.095","article-title":"A novel method for the quantification of adeno-associated virus vectors for RNA interference applications using quantitative polymerase chain reaction and purified genomic adeno-associated virus DNA as a standard","volume":"24","author":"Wagner","year":"2013","journal-title":"Hum. Gene Ther. Methods"}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/19\/10\/3129\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T15:25:10Z","timestamp":1760196310000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/19\/10\/3129"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2018,10,12]]},"references-count":69,"journal-issue":{"issue":"10","published-online":{"date-parts":[[2018,10]]}},"alternative-id":["ijms19103129"],"URL":"https:\/\/doi.org\/10.3390\/ijms19103129","relation":{},"ISSN":["1422-0067"],"issn-type":[{"value":"1422-0067","type":"electronic"}],"subject":[],"published":{"date-parts":[[2018,10,12]]}}}