Explanted diseased livers - a possible source of metabolic competent primary human hepatocytes
Being an integral part of basic, translational and clinical research, the demand for primary human hepatocytes (PHH) is continuously growing while the availability of tissue resection material for the isolation of metabolically competent PHH remains limited. To overcome current shortcomings, this st...
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| Veröffentlicht in: | PloS one 2014-07, Vol.9 (7), p.e101386 |
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| creator | Kleine, Moritz Riemer, Marc Krech, Till DeTemple, Daphne Jäger, Mark D Lehner, Frank Manns, Michael P Klempnauer, Jürgen Borlak, Jürgen Bektas, Hueseyin Vondran, Florian W R |
| description | Being an integral part of basic, translational and clinical research, the demand for primary human hepatocytes (PHH) is continuously growing while the availability of tissue resection material for the isolation of metabolically competent PHH remains limited. To overcome current shortcomings, this study evaluated the use of explanted diseased organs from liver transplantation patients as a potential source of PHH. Therefore, PHH were isolated from resected surgical specimens (Rx-group; n = 60) and explanted diseased livers obtained from graft recipients with low labMELD-score (Ex-group; n = 5). Using established protocols PHH were subsequently cultured for a period of 7 days. The viability and metabolic competence of cultured PHH was assessed by the following parameters: morphology and cell count (CyQuant assay), albumin synthesis, urea production, AST-leakage, and phase I and II metabolism. Both groups were compared in terms of cell yield and metabolic function, and results were correlated with clinical parameters of tissue donors. Notably, cellular yields and viabilities were comparable between the Rx- and Ex-group and were 5.3±0.5 and 2.9±0.7×106 cells/g liver tissue with 84.3±1.3 and 76.0±8.6% viability, respectively. Moreover, PHH isolated from the Rx- or Ex-group did not differ in regards to loss of cell number in culture, albumin synthesis, urea production, AST-leakage, and phase I and II metabolism (measured by the 7-ethoxycoumarin-O-deethylase and uracil-5'-diphosphate-glucuronyltransferase activity). Likewise, basal transcript expressions of the CYP monooxygenases 1A1, 2C8 and 3A4 were comparable as was their induction when treated with a cocktail that consisted of 3-methylcholantren, rifampicin and phenobarbital, with increased expression of CYP 1A1 and 3A4 mRNA while transcript expression of CYP 2C8 was only marginally changed. In conclusion, the use of explanted diseased livers obtained from recipients with low labMELD-score might represent a valuable source of metabolically competent PHH which are comparable in viability and function to cells obtained from specimens following partial liver resection. |
| doi_str_mv | 10.1371/journal.pone.0101386 |
| format | Article |
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To overcome current shortcomings, this study evaluated the use of explanted diseased organs from liver transplantation patients as a potential source of PHH. Therefore, PHH were isolated from resected surgical specimens (Rx-group; n = 60) and explanted diseased livers obtained from graft recipients with low labMELD-score (Ex-group; n = 5). Using established protocols PHH were subsequently cultured for a period of 7 days. The viability and metabolic competence of cultured PHH was assessed by the following parameters: morphology and cell count (CyQuant assay), albumin synthesis, urea production, AST-leakage, and phase I and II metabolism. Both groups were compared in terms of cell yield and metabolic function, and results were correlated with clinical parameters of tissue donors. Notably, cellular yields and viabilities were comparable between the Rx- and Ex-group and were 5.3±0.5 and 2.9±0.7×106 cells/g liver tissue with 84.3±1.3 and 76.0±8.6% viability, respectively. Moreover, PHH isolated from the Rx- or Ex-group did not differ in regards to loss of cell number in culture, albumin synthesis, urea production, AST-leakage, and phase I and II metabolism (measured by the 7-ethoxycoumarin-O-deethylase and uracil-5'-diphosphate-glucuronyltransferase activity). Likewise, basal transcript expressions of the CYP monooxygenases 1A1, 2C8 and 3A4 were comparable as was their induction when treated with a cocktail that consisted of 3-methylcholantren, rifampicin and phenobarbital, with increased expression of CYP 1A1 and 3A4 mRNA while transcript expression of CYP 2C8 was only marginally changed. In conclusion, the use of explanted diseased livers obtained from recipients with low labMELD-score might represent a valuable source of metabolically competent PHH which are comparable in viability and function to cells obtained from specimens following partial liver resection.</description><identifier>ISSN: 1932-6203</identifier><identifier>EISSN: 1932-6203</identifier><identifier>DOI: 10.1371/journal.pone.0101386</identifier><identifier>PMID: 24999631</identifier><language>eng</language><publisher>United States: Public Library of Science</publisher><subject>Adult ; Aged ; Albumin ; Albumins - biosynthesis ; Albumins - genetics ; Aryl Hydrocarbon Hydroxylases - genetics ; Cell Count ; Cell culture ; Cell number ; Cell Survival ; Cytology ; Female ; Gastroenterology ; Gene expression ; Gene Expression Regulation ; Hepatocytes ; Hepatocytes - metabolism ; Hepatocytes - pathology ; Hepatology ; Humans ; Infections ; Laparoscopy ; Leakage ; Liver ; Liver - pathology ; Liver - surgery ; Liver diseases ; Liver Diseases - pathology ; Liver Transplantation ; Liver transplants ; Male ; Medical schools ; Medicine and Health Sciences ; Metabolism ; Middle Aged ; Organs ; Parameters ; PCB ; Pharmacology ; Phenobarbital ; Polychlorinated biphenyls ; Rifampin ; RNA, Messenger - genetics ; RNA, Messenger - metabolism ; Surgery ; Synthesis ; Toxicology ; Transcription ; Transplantation ; Transplants & implants ; Uracil ; Urea ; Urea - metabolism ; Viability ; Young Adult</subject><ispartof>PloS one, 2014-07, Vol.9 (7), p.e101386</ispartof><rights>2014 Kleine et al. This is an open-access article distributed under the terms of the Creative Commons Attribution License: http://creativecommons.org/licenses/by/4.0/ (the “License”), which permits unrestricted use, distribution, and reproduction in any medium, provided the original author and source are credited. Notwithstanding the ProQuest Terms and Conditions, you may use this content in accordance with the terms of the License.</rights><rights>2014 Kleine et al 2014 Kleine et al</rights><lds50>peer_reviewed</lds50><oa>free_for_read</oa><woscitedreferencessubscribed>false</woscitedreferencessubscribed><citedby>FETCH-LOGICAL-c526t-85a96985d2cc599e35b5620dbab319c6e6422393a4a330d42f6926073cb92b223</citedby><cites>FETCH-LOGICAL-c526t-85a96985d2cc599e35b5620dbab319c6e6422393a4a330d42f6926073cb92b223</cites></display><links><openurl>$$Topenurl_article</openurl><openurlfulltext>$$Topenurlfull_article</openurlfulltext><thumbnail>$$Tsyndetics_thumb_exl</thumbnail><linktopdf>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4084809/pdf/$$EPDF$$P50$$Gpubmedcentral$$Hfree_for_read</linktopdf><linktohtml>$$Uhttps://www.ncbi.nlm.nih.gov/pmc/articles/PMC4084809/$$EHTML$$P50$$Gpubmedcentral$$Hfree_for_read</linktohtml><link.rule.ids>230,315,729,782,786,866,887,2104,2930,23873,27931,27932,53798,53800</link.rule.ids><backlink>$$Uhttps://www.ncbi.nlm.nih.gov/pubmed/24999631$$D View this record in MEDLINE/PubMed$$Hfree_for_read</backlink></links><search><contributor>Starkel, Peter</contributor><creatorcontrib>Kleine, Moritz</creatorcontrib><creatorcontrib>Riemer, Marc</creatorcontrib><creatorcontrib>Krech, Till</creatorcontrib><creatorcontrib>DeTemple, Daphne</creatorcontrib><creatorcontrib>Jäger, Mark D</creatorcontrib><creatorcontrib>Lehner, Frank</creatorcontrib><creatorcontrib>Manns, Michael P</creatorcontrib><creatorcontrib>Klempnauer, Jürgen</creatorcontrib><creatorcontrib>Borlak, Jürgen</creatorcontrib><creatorcontrib>Bektas, Hueseyin</creatorcontrib><creatorcontrib>Vondran, Florian W R</creatorcontrib><title>Explanted diseased livers - a possible source of metabolic competent primary human hepatocytes</title><title>PloS one</title><addtitle>PLoS One</addtitle><description>Being an integral part of basic, translational and clinical research, the demand for primary human hepatocytes (PHH) is continuously growing while the availability of tissue resection material for the isolation of metabolically competent PHH remains limited. 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a possible source of metabolic competent primary human hepatocytes</atitle><jtitle>PloS one</jtitle><addtitle>PLoS One</addtitle><date>2014-07-07</date><risdate>2014</risdate><volume>9</volume><issue>7</issue><spage>e101386</spage><pages>e101386-</pages><issn>1932-6203</issn><eissn>1932-6203</eissn><abstract>Being an integral part of basic, translational and clinical research, the demand for primary human hepatocytes (PHH) is continuously growing while the availability of tissue resection material for the isolation of metabolically competent PHH remains limited. To overcome current shortcomings, this study evaluated the use of explanted diseased organs from liver transplantation patients as a potential source of PHH. Therefore, PHH were isolated from resected surgical specimens (Rx-group; n = 60) and explanted diseased livers obtained from graft recipients with low labMELD-score (Ex-group; n = 5). Using established protocols PHH were subsequently cultured for a period of 7 days. The viability and metabolic competence of cultured PHH was assessed by the following parameters: morphology and cell count (CyQuant assay), albumin synthesis, urea production, AST-leakage, and phase I and II metabolism. Both groups were compared in terms of cell yield and metabolic function, and results were correlated with clinical parameters of tissue donors. Notably, cellular yields and viabilities were comparable between the Rx- and Ex-group and were 5.3±0.5 and 2.9±0.7×106 cells/g liver tissue with 84.3±1.3 and 76.0±8.6% viability, respectively. Moreover, PHH isolated from the Rx- or Ex-group did not differ in regards to loss of cell number in culture, albumin synthesis, urea production, AST-leakage, and phase I and II metabolism (measured by the 7-ethoxycoumarin-O-deethylase and uracil-5'-diphosphate-glucuronyltransferase activity). Likewise, basal transcript expressions of the CYP monooxygenases 1A1, 2C8 and 3A4 were comparable as was their induction when treated with a cocktail that consisted of 3-methylcholantren, rifampicin and phenobarbital, with increased expression of CYP 1A1 and 3A4 mRNA while transcript expression of CYP 2C8 was only marginally changed. In conclusion, the use of explanted diseased livers obtained from recipients with low labMELD-score might represent a valuable source of metabolically competent PHH which are comparable in viability and function to cells obtained from specimens following partial liver resection.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>24999631</pmid><doi>10.1371/journal.pone.0101386</doi><oa>free_for_read</oa></addata></record> |
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| issn | 1932-6203 1932-6203 |
| language | eng |
| recordid | cdi_plos_journals_2013276136 |
| source | MEDLINE; DOAJ Directory of Open Access Journals; Elektronische Zeitschriftenbibliothek - Frei zugängliche E-Journals; Public Library of Science (PLoS) Journals Open Access; PubMed Central; Free Full-Text Journals in Chemistry |
| subjects | Adult Aged Albumin Albumins - biosynthesis Albumins - genetics Aryl Hydrocarbon Hydroxylases - genetics Cell Count Cell culture Cell number Cell Survival Cytology Female Gastroenterology Gene expression Gene Expression Regulation Hepatocytes Hepatocytes - metabolism Hepatocytes - pathology Hepatology Humans Infections Laparoscopy Leakage Liver Liver - pathology Liver - surgery Liver diseases Liver Diseases - pathology Liver Transplantation Liver transplants Male Medical schools Medicine and Health Sciences Metabolism Middle Aged Organs Parameters PCB Pharmacology Phenobarbital Polychlorinated biphenyls Rifampin RNA, Messenger - genetics RNA, Messenger - metabolism Surgery Synthesis Toxicology Transcription Transplantation Transplants & implants Uracil Urea Urea - metabolism Viability Young Adult |
| title | Explanted diseased livers - a possible source of metabolic competent primary human hepatocytes |
| url | https://sfx.bib-bvb.de/sfx_tum?ctx_ver=Z39.88-2004&ctx_enc=info:ofi/enc:UTF-8&ctx_tim=2024-12-04T21%3A53%3A52IST&url_ver=Z39.88-2004&url_ctx_fmt=infofi/fmt:kev:mtx:ctx&rfr_id=info:sid/primo.exlibrisgroup.com:primo3-Article-proquest_plos_&rft_val_fmt=info:ofi/fmt:kev:mtx:journal&rft.genre=article&rft.atitle=Explanted%20diseased%20livers%20-%20a%20possible%20source%20of%20metabolic%20competent%20primary%20human%20hepatocytes&rft.jtitle=PloS%20one&rft.au=Kleine,%20Moritz&rft.date=2014-07-07&rft.volume=9&rft.issue=7&rft.spage=e101386&rft.pages=e101386-&rft.issn=1932-6203&rft.eissn=1932-6203&rft_id=info:doi/10.1371/journal.pone.0101386&rft_dat=%3Cproquest_plos_%3E2013276136%3C/proquest_plos_%3E%3Curl%3E%3C/url%3E&disable_directlink=true&sfx.directlink=off&sfx.report_link=0&rft_id=info:oai/&rft_pqid=2013276136&rft_id=info:pmid/24999631&rft_doaj_id=oai_doaj_org_article_e9b7054514af451dbb912c3101cd420c&rfr_iscdi=true |