{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,4,10]],"date-time":"2026-04-10T11:08:35Z","timestamp":1775819315501,"version":"3.50.1"},"reference-count":34,"publisher":"Wiley","issue":"1","license":[{"start":{"date-parts":[[2019,8,21]],"date-time":"2019-08-21T00:00:00Z","timestamp":1566345600000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81670593"],"award-info":[{"award-number":["81670593"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81870446"],"award-info":[{"award-number":["81870446"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81671838"],"award-info":[{"award-number":["81671838"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]},{"DOI":"10.13039\/501100001809","name":"National Natural Science Foundation of China","doi-asserted-by":"publisher","award":["81900571"],"award-info":[{"award-number":["81900571"]}],"id":[{"id":"10.13039\/501100001809","id-type":"DOI","asserted-by":"publisher"}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["Xenotransplantation"],"published-print":{"date-parts":[[2020,1]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>In vivo pig liver xenotransplantation preclinical trials appear to have poor efficiency compared to heart or kidney xenotransplantation because of xenogeneic rejection, including coagulopathy, and particularly thrombocytopenia. In contrast, ex vivo pig liver (wild type) perfusion systems have been proven to be effective in \u201cbridging\u201d liver failure patients until subsequent liver allotransplantation, and transgenic (human CD55\/CD59) modifications have even prolonged the duration of pig liver perfusion. Despite the fact that hepatocyte cell lines have also been proposed for extracorporeal blood circulation in conditions of acute liver failure, porcine hepatocyte cell lines, and the GalT\u2010KO background in particular, have not been developed and applied in this field. Herein, we established immortalized wild\u2010type and GalT\u2010KO porcine hepatocyte cell lines, which can be used for artificial liver support systems, cell transplantation, and even in vitro studies of xenotransplantation.<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Primary hepatocytes extracted from GalT\u2010KO and wild\u2010type pigs were transfected with SV40 LT lentivirus to establish immortalized GalT\u2010KO porcine hepatocytes (GalT\u2010KO\u2010hep) and wild\u2010type porcine hepatocytes (WT). Hepatocyte biomarkers and function\u2010related genes were assessed by immunofluorescence, periodic acid\u2010Schiff staining, indocyanine green (ICG) uptake, biochemical analysis, ELISA, and RT\u2010PCR. Furthermore, the tumorigenicity of immortalized cells was detected. In addition, a complement\u2010dependent cytotoxicity (CDC) assay was performed with GalT\u2010KO\u2010hep and WT cells. Cell death and viability rates were assessed by flow cytometry and CCK\u20108 assay.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>GalT\u2010KO and wild\u2010type porcine hepatocytes were successfully immortalized and maintained the characteristics of primary porcine hepatocytes, including albumin secretion, ICG uptake, urea and glycogen production, and expression of hepatocyte marker proteins and specific metabolic enzymes. GalT\u2010KO\u2010hep and WT cells were confirmed as having no tumorigenicity. In addition, GalT\u2010KO\u2010hep cells showed less apoptosis and more viability than WT cells when exposed to complement and xenogeneic serum.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusions<\/jats:title><jats:p>Two types of immortalized cell lines of porcine hepatocytes with GalT\u2010KO and wild\u2010type backgrounds were successfully established. GalT\u2010KO\u2010hep cells exhibited higher viability and injury resistance against a xenogeneic immune response.<\/jats:p><\/jats:sec>","DOI":"10.1111\/xen.12550","type":"journal-article","created":{"date-parts":[[2019,8,22]],"date-time":"2019-08-22T06:14:54Z","timestamp":1566454494000},"update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":3,"title":["Immortalization of porcine hepatocytes with a \u03b1\u20101,3\u2010galactosyltransferase knockout background"],"prefix":"10.1111","volume":"27","author":[{"given":"Quancheng","family":"Wang","sequence":"first","affiliation":[{"name":"Department of Hepatobiliary Surgery, Xijing Hospital The Fourth Military Medical University Xi'an China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0003-2937-9141","authenticated-orcid":false,"given":"Xuan","family":"Zhang","sequence":"additional","affiliation":[{"name":"Department of Hepatobiliary Surgery, Xijing Hospital The Fourth Military Medical University Xi'an China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Bo","family":"Wang","sequence":"additional","affiliation":[{"name":"Department of Hepatobiliary Surgery, Xijing Hospital The Fourth Military Medical University Xi'an China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Ge","family":"Bai","sequence":"additional","affiliation":[{"name":"Department of Hepatobiliary Surgery, Xijing Hospital The Fourth Military Medical University Xi'an China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-7994-1465","authenticated-orcid":false,"given":"Dengke","family":"Pan","sequence":"additional","affiliation":[{"name":"Organ Transplant and Clinical Immunology Translational Medicine Key Laboratory of Sichuan Province Sichuan Academy of an Transplant Science &amp; Sichuan Provincial People's Hospital Chengdu China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Peijun","family":"Yang","sequence":"additional","affiliation":[{"name":"Department of Hepatobiliary Surgery, Xijing Hospital The Fourth Military Medical University Xi'an China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Kaishan","family":"Tao","sequence":"additional","affiliation":[{"name":"Department of Hepatobiliary Surgery, Xijing Hospital The Fourth Military Medical University Xi'an China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Xiao","family":"Li","sequence":"additional","affiliation":[{"name":"Department of Hepatobiliary Surgery, Xijing Hospital The Fourth Military Medical University Xi'an China"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0002-8857-5663","authenticated-orcid":false,"given":"Kefeng","family":"Dou","sequence":"additional","affiliation":[{"name":"Department of Hepatobiliary Surgery, Xijing Hospital The Fourth Military Medical University Xi'an China"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2019,8,21]]},"reference":[{"key":"e_1_2_8_2_1","doi-asserted-by":"publisher","DOI":"10.1111\/jocn.14154"},{"issue":"473","key":"e_1_2_8_3_1","first-page":"e415","article-title":"Interspecies chimerism with mammalian pluripotent stem cells","volume":"168","author":"Wu J","year":"2017","journal-title":"Cell"},{"key":"e_1_2_8_4_1","doi-asserted-by":"publisher","DOI":"10.1111\/xen.12431"},{"key":"e_1_2_8_5_1","doi-asserted-by":"publisher","DOI":"10.1111\/xen.12497"},{"key":"e_1_2_8_6_1","doi-asserted-by":"publisher","DOI":"10.1038\/s41586-018-0765-z"},{"key":"e_1_2_8_7_1","doi-asserted-by":"publisher","DOI":"10.1097\/SLA.0000000000002977"},{"key":"e_1_2_8_8_1","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM199407283310404"},{"key":"e_1_2_8_9_1","doi-asserted-by":"publisher","DOI":"10.1097\/00007890-200001270-00013"},{"key":"e_1_2_8_10_1","doi-asserted-by":"publisher","DOI":"10.1034\/j.1399-3089.2002.1o058.x"},{"key":"e_1_2_8_11_1","doi-asserted-by":"publisher","DOI":"10.1371\/journal.pone.0175529"},{"key":"e_1_2_8_12_1","doi-asserted-by":"publisher","DOI":"10.1186\/s13613-018-0453-z"},{"key":"e_1_2_8_13_1","doi-asserted-by":"publisher","DOI":"10.1002\/hep.1840040332"},{"key":"e_1_2_8_14_1","doi-asserted-by":"publisher","DOI":"10.1097\/01.MAT.0000161045.16805.8B"},{"key":"e_1_2_8_15_1","first-page":"7","article-title":"Establishment of a tightly regulated human cell line for the development of hepatocyte transplantation","volume":"13","author":"Kobayashi N","year":"2000","journal-title":"Hum Cell"},{"key":"e_1_2_8_16_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.90.11.5123"},{"key":"e_1_2_8_17_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.transproceed.2009.11.043"},{"key":"e_1_2_8_18_1","doi-asserted-by":"publisher","DOI":"10.3748\/wjg.v16.i13.1660"},{"key":"e_1_2_8_19_1","doi-asserted-by":"publisher","DOI":"10.3727\/096368908786991542"},{"key":"e_1_2_8_20_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jhep.2007.02.019"},{"key":"e_1_2_8_21_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jceh.2014.08.001"},{"key":"e_1_2_8_22_1","doi-asserted-by":"publisher","DOI":"10.1111\/xen.12492"},{"key":"e_1_2_8_23_1","doi-asserted-by":"publisher","DOI":"10.1016\/S0091-679X(08)61797-5"},{"key":"e_1_2_8_24_1","doi-asserted-by":"publisher","DOI":"10.1006\/excr.1998.4272"},{"key":"e_1_2_8_25_1","doi-asserted-by":"publisher","DOI":"10.4049\/jimmunol.174.11.7302"},{"key":"e_1_2_8_26_1","doi-asserted-by":"publisher","DOI":"10.1002\/cbin.10278"},{"key":"e_1_2_8_27_1","doi-asserted-by":"publisher","DOI":"10.1007\/s00441-005-0101-4"},{"key":"e_1_2_8_28_1","doi-asserted-by":"publisher","DOI":"10.1006\/mthe.2000.0141"},{"key":"e_1_2_8_29_1","doi-asserted-by":"publisher","DOI":"10.1097\/00007890-199312000-00031"},{"key":"e_1_2_8_30_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.jhep.2004.11.041"},{"key":"e_1_2_8_31_1","doi-asserted-by":"publisher","DOI":"10.1084\/jem.160.5.1519"},{"key":"e_1_2_8_32_1","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.84.5.1369"},{"key":"e_1_2_8_33_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.ijsu.2015.08.077"},{"key":"e_1_2_8_34_1","doi-asserted-by":"publisher","DOI":"10.1016\/j.transproceed.2017.03.090"},{"key":"e_1_2_8_35_1","doi-asserted-by":"publisher","DOI":"10.3727\/096368916X691286"}],"container-title":["Xenotransplantation"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1111%2Fxen.12550","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/xen.12550","content-type":"application\/pdf","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/full-xml\/10.1111\/xen.12550","content-type":"application\/xml","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1111\/xen.12550","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,9,8]],"date-time":"2023-09-08T10:05:30Z","timestamp":1694167530000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1111\/xen.12550"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2019,8,21]]},"references-count":34,"journal-issue":{"issue":"1","published-print":{"date-parts":[[2020,1]]}},"alternative-id":["10.1111\/xen.12550"],"URL":"https:\/\/doi.org\/10.1111\/xen.12550","archive":["Portico"],"relation":{},"ISSN":["0908-665X","1399-3089"],"issn-type":[{"value":"0908-665X","type":"print"},{"value":"1399-3089","type":"electronic"}],"subject":[],"published":{"date-parts":[[2019,8,21]]},"assertion":[{"value":"2019-02-20","order":0,"name":"received","label":"Received","group":{"name":"publication_history","label":"Publication History"}},{"value":"2019-07-24","order":1,"name":"accepted","label":"Accepted","group":{"name":"publication_history","label":"Publication History"}},{"value":"2019-08-21","order":2,"name":"published","label":"Published","group":{"name":"publication_history","label":"Publication History"}}],"article-number":"e12550"}}