{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,1,6]],"date-time":"2026-01-06T13:42:37Z","timestamp":1767706957977,"version":"build-2065373602"},"reference-count":59,"publisher":"MDPI AG","issue":"2","license":[{"start":{"date-parts":[[2023,2,20]],"date-time":"2023-02-20T00:00:00Z","timestamp":1676851200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"Wellington Research Fellowship"},{"name":"National Institute for Health Research (NIHR) UCLH\/UCL Biomedical Research Centre (BRC)"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Bioengineering"],"abstract":"<jats:p>Introduction: Allogenic hepatocyte transplantation is an attractive alternative to whole-organ transplantation, particularly for the treatment of metabolic disorders and acute liver failure. However, the shortage of human donor organs for cell isolation, the low cell yield from decellularisation regimes, and low engraftment rates from portal administration of donor cells have restricted its clinical application. Using ultrasound histotripsy to provide a nidus in the liver for direct cell transplantation offers a new approach to overcoming key limitations in current cell therapy. We have analysed the liver cavity constituents to assess their potential as a site for cell delivery and implantation. Methods: Using human organ retrieval techniques, pig livers were collected from the abattoir and transported in ice-cold storage to the laboratory. Following 2 h of cold storage, the livers were flushed with organ preservation solution and placed on an organ perfusion circuit to maintain viability. Organs were perfused with Soltran\u2122 organ preservation solution via the portal vein at a temperature of 24\u201330 \u00b0C. The perfusion circuit was oxygenated through equilibration with room air. Perfused livers (n=5) were subjected to ultrasound histotripsy, producing a total of 130 lesions. Lesions were generated by applying 50 pulses at 1 Hz pulse repetition frequency and 1% duty cycle using a single element 2 MHz bowl-shaped transducer (Sonic Concepts, H-148). Following histotripsy, a focal liver lesion was produced, which had a liquid centre. The fluid from each lesion was aspirated and cultured in medium (RPMI) at 37 \u00b0C in an incubator. Cell cultures were analysed at 1 and 7 days for cell viability and a live-dead assay was performed. The histotripsy sites were excised following aspiration and H&amp;E staining was used to characterise the liver lesions. Cell morphology was determined by histology. Results: Histotripsy created a subcapsular lesion (~5 mm below the liver capsule; size ranging from 3 to 5 mm), which contained a suspension of cells. On average, 61\u00d7104 cells per mL were isolated. Hepatocytes were present in the aspirate, were viable at 24 h post isolation and remained viable in culture for up to 1 week, as determined by phalloidin\/DAPI cell viability stains. Cultures up to 21 days revealed metabolically active live hepatocyte. Live-dead assays confirmed hepatocyte viability at 1 week (Day 1: 12% to Day 7: 45% live cells; p &lt; 0.0001), which retained metabolic activity and morphology, confirmed on assay and microscopy. Cell Titre-GloTM showed a peak metabolic activity at 1 week (average luminescence 24.6 RLU; p &lt; 0.0001) post-culture compared with the control (culture medium alone), reduced to 1\/3 of peak level (7.85 RLU) by day 21. Conclusions: Histotripsy of the liver allows isolation and culture of hepatocytes with a high rate of viability after 1 week in culture. Reproducing these findings using human livers may lead to wide clinical applications in cell therapy.<\/jats:p>","DOI":"10.3390\/bioengineering10020276","type":"journal-article","created":{"date-parts":[[2023,2,21]],"date-time":"2023-02-21T01:39:28Z","timestamp":1676943568000},"page":"276","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":6,"title":["Liver Ultrasound Histotripsy: Novel Analysis of the Histotripsy Site Cell Constituents with Implications for Histotripsy Application in Cell Transplantation and Cancer Therapy"],"prefix":"10.3390","volume":"10","author":[{"given":"Saied","family":"Froghi","sequence":"first","affiliation":[{"name":"Department of HPB & Liver Transplantation Surgery, Royal Free London NHS Foundation Trust, Pond Street, Hampstead, London NW3 2QG, UK"},{"name":"Centre for Surgical Innovation, Organ Regeneration and Transplantation, UCL Division of Surgery & Interventional Sciences, Royal Free Hospital Campus, Pond Street, Hampstead, London NW3 2QG, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5682-0460","authenticated-orcid":false,"given":"Matheus Oliveira","family":"de Andrade","sequence":"additional","affiliation":[{"name":"Ultrasonics Group, Department of Mechanical Engineering, Roberts Engineering Building, University College London, Torrington Place, London WC1E 7JE, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Layla Mohammad","family":"Hadi","sequence":"additional","affiliation":[{"name":"Centre for Surgical Innovation, Organ Regeneration and Transplantation, UCL Division of Surgery & Interventional Sciences, Royal Free Hospital Campus, Pond Street, Hampstead, London NW3 2QG, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pierre","family":"Gelat","sequence":"additional","affiliation":[{"name":"Centre for Surgical Innovation, Organ Regeneration and Transplantation, UCL Division of Surgery & Interventional Sciences, Royal Free Hospital Campus, Pond Street, Hampstead, London NW3 2QG, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-8078-6688","authenticated-orcid":false,"given":"Hassan","family":"Rashidi","sequence":"additional","affiliation":[{"name":"Stem Cell & Regenerative Medicine Section, UCL Great Ormond Street Institute of Child Health, London WC1N 1EH, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Alberto","family":"Quaglia","sequence":"additional","affiliation":[{"name":"Department of Cellular Pathology, Royal Free London NHS Foundation Trust, Pond Street, Hampstead, London NW3 2QG, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5752-5295","authenticated-orcid":false,"given":"Barry","family":"Fuller","sequence":"additional","affiliation":[{"name":"Centre for Surgical Innovation, Organ Regeneration and Transplantation, UCL Division of Surgery & Interventional Sciences, Royal Free Hospital Campus, Pond Street, Hampstead, London NW3 2QG, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Nader","family":"Saffari","sequence":"additional","affiliation":[{"name":"Ultrasonics Group, Department of Mechanical Engineering, Roberts Engineering Building, University College London, Torrington Place, London WC1E 7JE, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Brian","family":"Davidson","sequence":"additional","affiliation":[{"name":"Department of HPB & Liver Transplantation Surgery, Royal Free London NHS Foundation Trust, Pond Street, Hampstead, London NW3 2QG, UK"},{"name":"Centre for Surgical Innovation, Organ Regeneration and Transplantation, UCL Division of Surgery & Interventional Sciences, Royal Free Hospital Campus, Pond Street, Hampstead, London NW3 2QG, UK"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,2,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"232","DOI":"10.1038\/pr.2017.284","article-title":"Human hepatocyte transplantation for liver disease: Current status and future perspectives","volume":"83","author":"Iansante","year":"2018","journal-title":"Pediatr. Res."},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"288","DOI":"10.1038\/nrgastro.2010.44","article-title":"Human hepatocyte transplantation: Current experience and future challenges","volume":"7","author":"Dhawan","year":"2010","journal-title":"Nat. Rev. Gastroenterol. Hepatol."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"713","DOI":"10.1002\/lt.20732","article-title":"Isolation of hepatocytes from livers from non-heart-beating donors for cell transplantation","volume":"12","author":"Hughes","year":"2006","journal-title":"Liver Transplant."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"298","DOI":"10.1067\/mlc.2001.119148","article-title":"Liver cell transplantation: The road to clinical application","volume":"138","author":"Allen","year":"2001","journal-title":"J. Lab. Clin. Med."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"431","DOI":"10.1007\/s10545-006-0245-8","article-title":"Hepatocyte transplantation for liver-based metabolic disorders","volume":"29","author":"Dhawan","year":"2006","journal-title":"J. Inherit. Metab. Dis."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"441","DOI":"10.1097\/01.tp.0000231689.44266.ac","article-title":"Human Hepatocyte Transplantation: Worldwide Results","volume":"82","author":"Fisher","year":"2006","journal-title":"Transplantation"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"1","DOI":"10.3727\/096368911X566208","article-title":"Improving the Techniques for Human Hepatocyte Transplantation: Report from a Consensus Meeting in London","volume":"21","author":"Puppi","year":"2012","journal-title":"Cell Transplant."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"S39","DOI":"10.1002\/lt.24226","article-title":"Clinical human hepatocyte transplantation: Current status and challenges","volume":"21","author":"Dhawan","year":"2015","journal-title":"Liver Transplant."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"463","DOI":"10.1016\/S1084952102001350","article-title":"Progress in human hepatocytes: Isolation, culture & cryopreservation","volume":"13","author":"Mitry","year":"2002","journal-title":"Semin. Cell Dev. Biol."},{"key":"ref_10","first-page":"e53069","article-title":"Protocol for Isolation of Primary Human Hepatocytes and Corresponding Major Populations of Non-parenchymal Liver Cells","volume":"109","author":"Kegel","year":"2016","journal-title":"J. Vis. Exp."},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1002\/lt.22467","article-title":"Cell therapies for liver diseases","volume":"18","author":"Yu","year":"2012","journal-title":"Liver Transplant."},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"235023","DOI":"10.1088\/1361-6560\/aaef16","article-title":"Mechanical decellularization of tissue volumes using boiling histotripsy","volume":"63","author":"Wang","year":"2018","journal-title":"Phys. Med. Biol."},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"145","DOI":"10.3109\/02656736.2015.1007538","article-title":"Histotripsy methods in mechanical disintegration of tissue: Towards clinical applications","volume":"31","author":"Khokhlova","year":"2015","journal-title":"Int. J. Hyperth."},{"key":"ref_14","doi-asserted-by":"crossref","unstructured":"Eranki, A., Farr, N., Partanen, A., Sharma, K.V., Chen, H., Rossi, C.T., Kothapalli, S.V.V.N., Oetgen, M., Kim, A., and Negussie, A.H. (2017). Boiling histotripsy lesion characterization on a clinical magnetic resonance imaging-guided high intensity focused ultrasound system. PLoS ONE, 12.","DOI":"10.1371\/journal.pone.0173867"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"262","DOI":"10.1016\/j.ultsonch.2019.01.019","article-title":"The effects of ultrasound pressure and temperature fields in millisecond bubble nucleation","volume":"55","author":"Haqshenas","year":"2019","journal-title":"Ultrason. Sonochem."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"164","DOI":"10.1016\/j.ultsonch.2019.01.001","article-title":"Mechanical damage induced by the appearance of rectified bubble growth in a viscoelastic medium during boiling histotripsy exposure","volume":"53","author":"Pahk","year":"2019","journal-title":"Ultrason. Sonochem."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"2848","DOI":"10.1016\/j.ultrasmedbio.2017.08.938","article-title":"Numerical and Experimental Study of Mechanisms Involved in Boiling Histotripsy","volume":"43","author":"Pahk","year":"2017","journal-title":"Ultrasound Med. Biol."},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"2673","DOI":"10.1016\/j.ultrasmedbio.2018.07.025","article-title":"Bubble dynamics in boiling histotripsy","volume":"44","author":"Pahk","year":"2018","journal-title":"Ultrasound Med. Biol."},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"1958","DOI":"10.1016\/j.ultrasmedbio.2016.03.020","article-title":"A Novel Approach to Ultrasound-Mediated Tissue Decellularization and Intra-Hepatic Cell Delivery in Rats","volume":"42","author":"Pahk","year":"2016","journal-title":"Ultrasound Med. Biol."},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"506","DOI":"10.1083\/jcb.43.3.506","article-title":"High-yield preparation of isolated rat liver parenchymal cells: A biochemical and fine structural study","volume":"43","author":"Berry","year":"1969","journal-title":"J. Cell Biol."},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"343","DOI":"10.1080\/02656736.2016.1255918","article-title":"Cavitation enhances coagulated size during pulsed high-intensity focussed ultrasound ablation in an isolated liver perfusion system","volume":"33","author":"Zhao","year":"2017","journal-title":"Int. J. Hyperth."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"165","DOI":"10.1063\/1.3131405","article-title":"A User-Friendly Software Package for HIFU Simulation","volume":"1113","author":"Soneson","year":"2009","journal-title":"AIP Conf. Proc."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3498","DOI":"10.1121\/1.3626152","article-title":"Controlled tissue emulsification produced by high intensity focused ultrasound shock waves and millisecond boiling","volume":"130","author":"Khokhlova","year":"2011","journal-title":"J. Acoust. Soc. Am."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"2406","DOI":"10.1121\/1.2967836","article-title":"Acoustic characterization of high intensity focused ultrasound fields: A combined measurement and modeling approach","volume":"124","author":"Canney","year":"2008","journal-title":"J. Acoust. Soc. Am."},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"615","DOI":"10.1038\/labinvest.2017.13","article-title":"Blue intensity matters for cell cycle profiling in fluorescence DAPI-stained images","volume":"97","author":"Ferro","year":"2017","journal-title":"Lab. Investig."},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"300","DOI":"10.1016\/j.bbcan.2016.10.002","article-title":"Guidelines for the selection of functional assays to evaluate the hallmarks of cancer","volume":"1866","author":"Sukumar","year":"2016","journal-title":"Biochim. Biophys. Acta (BBA)\u2014Rev. Cancer"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"187","DOI":"10.1016\/j.hpb.2020.06.013","article-title":"Systematic review of the role of high intensity focused ultrasound (HIFU) in treating malignant lesions of the hepatobiliary system","volume":"23","author":"Sehmbi","year":"2021","journal-title":"HPB"},{"key":"ref_28","doi-asserted-by":"crossref","unstructured":"Worlikar, T., Vlaisavljevich, E., Gerhardson, T., Greve, J., Wan, S., Kuruvilla, S., Lundt, J., Ives, K., Hall, T., and Welling, T.H. (2018, January 18\u201321). Histotripsy for Non-Invasive Ablation of Hepatocellular Carcinoma (HCC) Tumor in a Subcutaneous Xenograft Murine Model. Proceedings of the 40th Annual International Conference of the IEEE Engineering in Medicine and Biology Society (EMBC), Honolulu, HI, USA.","DOI":"10.1109\/EMBC.2018.8513650"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"331","DOI":"10.1109\/TBME.2022.3191069","article-title":"Histotripsy Ablation of Spontaneously Occurring Canine Bone Tumors","volume":"70","author":"Ruger","year":"2023","journal-title":"IEEE Trans. Biomed. Eng."},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"E504","DOI":"10.1149\/2.1031810jes","article-title":"Explanation of Bubble Nucleation Mechanisms: A Gradient Theory Approach","volume":"165","author":"Vachaparambil","year":"2018","journal-title":"J. Electrochem. Soc."},{"key":"ref_31","doi-asserted-by":"crossref","first-page":"1237","DOI":"10.1016\/j.ultrasmedbio.2017.01.016","article-title":"Non-Invasive Liver Ablation Using Histotripsy: Preclinical Safety Study in an In Vivo Porcine Model","volume":"43","author":"Vlaisavljevich","year":"2017","journal-title":"Ultrasound Med. Biol."},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"20176","DOI":"10.1038\/s41598-019-56658-7","article-title":"Pilot in vivo studies on transcutaneous boiling histotripsy in porcine liver and kidney","volume":"9","author":"Khokhlova","year":"2019","journal-title":"Sci. Rep."},{"key":"ref_33","doi-asserted-by":"crossref","unstructured":"Panconesi, R., Flores Carvalho, M., Mueller, M., Meierhofer, D., Dutkowski, P., Muiesan, P., and Schlegel, A. (2021). Viability Assessment in Liver Transplantation\u2014What Is the Impact of Dynamic Organ Preservation?. Biomedicines, 9.","DOI":"10.3390\/biomedicines9020161"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"1890","DOI":"10.1016\/j.ultrasmedbio.2016.03.018","article-title":"Non-Invasive Ultrasound Liver Ablation Using Histotripsy: Chronic Study in an In Vivo Rodent Model","volume":"42","author":"Vlaisavljevich","year":"2016","journal-title":"Ultrasound Med. Biol."},{"key":"ref_35","doi-asserted-by":"crossref","unstructured":"Vlaisavljevich, E., Cain, C.A., and Xu, Z. (2011, January 18\u201321). The effect of histotripsy on tissues with different mechanical properties. Proceedings of the 2011 IEEE International Ultrasonics Symposium, Orlando, FL, USA.","DOI":"10.1109\/ULTSYM.2011.0369"},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"253","DOI":"10.1088\/0031-9155\/59\/2\/253","article-title":"Effects of tissue mechanical properties on susceptibility to histotripsy-induced tissue damage","volume":"59","author":"Vlaisavljevich","year":"2014","journal-title":"Phys. Med. Biol."},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"1485","DOI":"10.1002\/hep.31474","article-title":"Long-Term Perfusion of the Liver Outside the Body: Warming Up for Ex Vivo Therapies?","volume":"72","author":"Schurink","year":"2020","journal-title":"Hepatology"},{"key":"ref_38","unstructured":"Hafez, T., and Fuller, B.J. (2007). Advances in Biopreservation, CRC Press."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1","DOI":"10.1083\/jcb.66.1.1","article-title":"Isolation and subfractionation on ficoll gradients of adult rat hepatocytes. Size, morphology, and biochemical characteristics of cell fractions","volume":"66","author":"Drochmans","year":"1975","journal-title":"J. Cell Biol."},{"key":"ref_40","first-page":"e50615","article-title":"Isolation of Human Hepatocytes by a Two-Step Collagenase Perfusion Procedure","volume":"79","author":"Lee","year":"2013","journal-title":"J. Vis. Exp."},{"key":"ref_41","doi-asserted-by":"crossref","first-page":"1615","DOI":"10.1002\/lt.24829","article-title":"A systematic review and meta-analysis of cold in situ perfusion and preservation of the hepatic allograft: Working toward a unified approach","volume":"23","author":"Hameed","year":"2017","journal-title":"Liver Transplant."},{"key":"ref_42","first-page":"e56993","article-title":"Purification of Hepatocytes and Sinusoidal Endothelial Cells from Mouse Liver Perfusion","volume":"132","author":"Cabral","year":"2018","journal-title":"J. Vis. Exp."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"345","DOI":"10.1089\/10763270152044206","article-title":"Liver Tissue Engineering: A Role for Co-culture Systems in Modifying Hepatocyte Function and Viability","volume":"7","author":"Bhandari","year":"2001","journal-title":"Tissue Eng."},{"key":"ref_44","doi-asserted-by":"crossref","unstructured":"Shulman, M., and Nahmias, Y. (2012). Long-Term Culture and Coculture of Primary Rat and Human Hepatocytes, Humana Press.","DOI":"10.1007\/978-1-62703-125-7_17"},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"1279","DOI":"10.1007\/s10616-018-0219-3","article-title":"Co-culture system of hepatocytes and endothelial cells: Two in vitro approaches for enhancing liver-specific functions of hepatocytes","volume":"70","author":"Wang","year":"2018","journal-title":"Cytotechnology"},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"1348","DOI":"10.3748\/wjg.v22.i4.1348","article-title":"Hepatic non-parenchymal cells: Master regulators of alcoholic liver disease?","volume":"22","author":"Seo","year":"2016","journal-title":"World J. Gastroenterol."},{"key":"ref_47","doi-asserted-by":"crossref","first-page":"205","DOI":"10.1007\/s11626-008-9166-1","article-title":"Maintaining Hepatocyte Differentiation in vitro through Co-Culture with Hepatic Stellate Cells","volume":"45","author":"Krause","year":"2009","journal-title":"In Vitro Cell. Dev. Biol. Anim."},{"key":"ref_48","doi-asserted-by":"crossref","first-page":"1883","DOI":"10.1096\/fasebj.13.14.1883","article-title":"Effect of cell\u2013cell interactions in preservation of cellular phenotype: Cocultivation of hepatocytes and nonparenchymal cells","volume":"13","author":"Bhatia","year":"1999","journal-title":"FASEB J."},{"key":"ref_49","doi-asserted-by":"crossref","first-page":"67","DOI":"10.1159\/000091096","article-title":"The Effect of Three-Dimensional Co-Culture of Hepatocytes and Hepatic Stellate Cells on Key Hepatocyte Functions in vitro","volume":"181","author":"Thomas","year":"2005","journal-title":"Cells Tissues Organs"},{"key":"ref_50","doi-asserted-by":"crossref","first-page":"7413","DOI":"10.3748\/wjg.v12.i46.7413","article-title":"Role of Kupffer cells in the pathogenesis of liver disease","volume":"12","author":"Kolios","year":"2006","journal-title":"World J. Gastroenterol."},{"key":"ref_51","doi-asserted-by":"crossref","first-page":"574276","DOI":"10.3389\/fimmu.2020.574276","article-title":"Immunomodulatory Role of the Extracellular Matrix within the Liver Disease Microenvironment","volume":"11","author":"McQuitty","year":"2020","journal-title":"Front. Immunol."},{"key":"ref_52","doi-asserted-by":"crossref","first-page":"474","DOI":"10.1016\/j.bbrc.2020.06.041","article-title":"Kupffer cells mediate the recruitment of hepatic stellate cells into the localized liver damage","volume":"529","author":"Abbas","year":"2020","journal-title":"Biochem. Biophys. Res. Commun."},{"key":"ref_53","doi-asserted-by":"crossref","first-page":"364","DOI":"10.3892\/mmr.2021.12003","article-title":"Interaction of non-parenchymal hepatocytes in the process of hepatic fibrosis (Review)","volume":"23","author":"Cheng","year":"2021","journal-title":"Mol. Med. Rep."},{"key":"ref_54","doi-asserted-by":"crossref","first-page":"534","DOI":"10.1089\/ten.tec.2018.0134","article-title":"Three-Dimensional Spheroid Primary Human Hepatocytes in Monoculture and Coculture with Nonparenchymal Cells","volume":"24","author":"Baze","year":"2018","journal-title":"Tissue Eng. Part C Methods"},{"key":"ref_55","doi-asserted-by":"crossref","first-page":"3117","DOI":"10.1007\/s00204-018-2280-2","article-title":"3D human liver tissue from pluripotent stem cells displays stable phenotype in vitro and supports compromised liver function in vivo","volume":"92","author":"Rashidi","year":"2018","journal-title":"Arch. Toxicol."},{"key":"ref_56","doi-asserted-by":"crossref","first-page":"1010","DOI":"10.1002\/hep.1840080506","article-title":"Cellular interactions promote tissue-specific function, biomatrix deposition and junctional communication of primary cultured hepatocytes","volume":"8","author":"Goulet","year":"1988","journal-title":"Hepatology"},{"key":"ref_57","doi-asserted-by":"crossref","first-page":"29","DOI":"10.3109\/15419061.2016.1151875","article-title":"Structure, Regulation and Function of Gap Junctions in Liver","volume":"22","author":"Willebrords","year":"2015","journal-title":"Cell Commun. Adhes."},{"key":"ref_58","doi-asserted-by":"crossref","first-page":"299","DOI":"10.1080\/10408440600599273","article-title":"Involvement of Cell Junctions in Hepatocyte Culture Functionality","volume":"36","author":"Vinken","year":"2006","journal-title":"Crit. Rev. Toxicol."},{"key":"ref_59","first-page":"e9411","article-title":"Comparative Anatomical Study between the Human and Swine Liver and Its Importance in Xenotransplantation","volume":"12","author":"Ntonas","year":"2020","journal-title":"Cureus"}],"container-title":["Bioengineering"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/2306-5354\/10\/2\/276\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T18:37:26Z","timestamp":1760121446000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/2306-5354\/10\/2\/276"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,2,20]]},"references-count":59,"journal-issue":{"issue":"2","published-online":{"date-parts":[[2023,2]]}},"alternative-id":["bioengineering10020276"],"URL":"https:\/\/doi.org\/10.3390\/bioengineering10020276","relation":{},"ISSN":["2306-5354"],"issn-type":[{"type":"electronic","value":"2306-5354"}],"subject":[],"published":{"date-parts":[[2023,2,20]]}}}