{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,9,27]],"date-time":"2026-09-27T18:20:37Z","timestamp":1790533237352,"version":"4.1.0"},"reference-count":29,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2019,4,10]],"date-time":"2019-04-10T00:00:00Z","timestamp":1554854400000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/100007048","name":"Mayo Foundation for Medical Education and Research","doi-asserted-by":"publisher","id":[{"id":"10.13039\/100007048","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/100000098","name":"NIH Clinical Center","doi-asserted-by":"publisher","award":["R01 DK106667"],"award-info":[{"award-number":["R01 DK106667"]}],"id":[{"id":"10.13039\/100000098","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Xenotransplantation"],"published-print":{"date-parts":[[2019,7]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background and aims<\/jats:title><jats:p>Cell\u2010based therapies for liver disease such as bioartificial liver rely on a large quantity and high quality of hepatocytes. Cold storage was previously shown to be a better way to preserve the viability and functionality of hepatocytes during transportation rather than freezing, but this was only proved at a lower density of rat hepatocytes spheroids. The purpose of this study was to optimize conditions for cold storage of high density of primary porcine hepatocyte spheroids.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Porcine hepatocytes were isolated by a three\u2010step perfusion method; hepatocyte spheroids were formed by a 24\u00a0hours rocked culture technique. Hepatocyte cell density was 5\u00a0\u00d7\u00a010<jats:sup>6<\/jats:sup>\/mL in 1000\u00a0mL spheroid forming medium. Spheroids were then maintained in rocked culture at 37\u00b0C (control condition) or cold stored at 4\u00b0C for 24, 48 or 72\u00a0hours in four different cold storage solutions: histidine\u2010tryptophan\u2010ketoglutarate (HTK) alone; HTK\u00a0+\u00a01\u00a0mM deferoxamine (DEF); HTK\u00a0+\u00a05\u00a0mM <jats:italic>N<\/jats:italic>\u2010acetyl\u2010L\u2010cysteine (NAC); and HTK\u00a0+\u00a01\u00a0mM DEF\u00a0+\u00a05\u00a0mM NAC. The viability, ammonia clearance, albumin production, gene expression, and functional activity of cytochrome P450 enzymes were measured after recovery from the cold storage.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>In this study, we observed that cold\u2010induced injury was reduced by the addition of the iron chelator. Viability of HTK\u00a0+\u00a0DEF group hepatocyte spheroids was increased compared with other cold storage groups (<jats:italic>P<\/jats:italic>\u00a0&lt;\u00a00.05). Performance metrics of porcine hepatocyte spheroids cold stored for 24\u00a0hours were similar to those in control conditions. The hepatocyte spheroids in control conditions started to lose their ability to clear ammonia while production of albumin was still active at 48 and 72\u00a0hours (<jats:italic>P<\/jats:italic>\u00a0&lt;\u00a00.05). In contrast, the viability and functionality of hepatocyte spheroids including ammonia clearance and albumin secretion were preserved in HTK\u00a0+\u00a0DEF group at both 48\u2010 and 72\u2010hour time points (<jats:italic>P<\/jats:italic>\u00a0&lt;\u00a00.05).<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>The beneficial effects of HTK supplemented with DEF were more obvious after cold storage of high density of porcine hepatocyte spheroids for 72\u00a0hours. The porcine hepatocyte spheroids were above the cutoff criteria for use in a spheroid\u2010based bioartificial liver.<\/jats:p><\/jats:sec>","DOI":"10.1111\/xen.12512","type":"journal-article","created":{"date-parts":[[2019,4,10]],"date-time":"2019-04-10T07:17:44Z","timestamp":1554880664000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":8,"title":["Cold storage of porcine hepatocyte spheroids for spheroid bioartificial liver"],"prefix":"10.1111","volume":"26","author":[{"ORCID":"https:\/\/orcid.org\/0000-0001-8661-4617","authenticated-orcid":false,"given":"Yi","family":"Li","sequence":"first","affiliation":[{"name":"Laboratory of Pathology, Key Laboratory of Transplant Engineering and Immunology, NHFPC, West China Hospital Sichuan University Chengdu China"},{"name":"Department of Surgery Mayo Clinic Rochester Minnesota"},{"name":"William J. von Liebig Center for Transplantation and Clinical Regeneration Mayo Clinic Rochester Minnesota"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Harvey S.","family":"Chen","sequence":"additional","affiliation":[{"name":"Department of Surgery Mayo Clinic Rochester Minnesota"},{"name":"William J. von Liebig Center for Transplantation and Clinical Regeneration Mayo Clinic Rochester Minnesota"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Mohammed","family":"Shaheen","sequence":"additional","affiliation":[{"name":"Department of Surgery Mayo Clinic Rochester Minnesota"},{"name":"William J. von Liebig Center for Transplantation and Clinical Regeneration Mayo Clinic Rochester Minnesota"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Dong Jin","family":"Joo","sequence":"additional","affiliation":[{"name":"Department of Surgery Mayo Clinic Rochester Minnesota"},{"name":"William J. von Liebig Center for Transplantation and Clinical Regeneration Mayo Clinic Rochester 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