{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,5,20]],"date-time":"2025-05-20T14:29:07Z","timestamp":1747751347441},"reference-count":48,"publisher":"Wiley","issue":"2","license":[{"start":{"date-parts":[[2003,12,15]],"date-time":"2003-12-15T00:00:00Z","timestamp":1071446400000},"content-version":"vor","delay-in-days":1019,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["Journal of Viral Hepatitis"],"published-print":{"date-parts":[[2001,3]]},"abstract":"<jats:p>Studies on the <jats:italic>in vitro<\/jats:italic> hepatitis C virus (HCV) infection are hampered by the lack of an appropriate system to culture permissive cells to be continuously infected with HCV. Trypsinization required for cell passage can lead to possible temporary loss of permissiveness for infection, whereas refreshment of the medium can result in loss of infectious particles necessary for perpetuation of the infection; it is therefore very difficult to maintain a continuous HCV infection in cell cultures. A new infection method was designed and evaluated in order to prevent these unfavourable circumstances. A cell line derived from the human hepatoblastoma cell line Hep G2 was grown in the extracapillary space of a haemodialysis cartridge, in the presence of a HCV\u2010positive inoculum, while the culture medium was recirculated through the intracapillary space, supplying the cells with nutrients and oxygen. HCV RNA could continuously be detected in the cells up to 77\u2003days of culture. Sequence analysis of the HCV hypervariable region 1 (HVR1) revealed that 56% and 75%, respectively, of the clones obtained from the cells at day 20 and 40 after start of the infection were different from the clones obtained from the original inoculum and that certain nucleotide positions in this region were more susceptible to mutations, leading to an alteration in amino acid sequence. As none of these sequences were present in the clones from the inoculum, it is suggested that new HCV quasispecies have emerged as a result of viral replication in the hepatocytes <jats:italic>in vitro<\/jats:italic>. This system seems a valuable tool for the <jats:italic>in vitro<\/jats:italic> evaluation of antiviral drugs.<\/jats:p>","DOI":"10.1046\/j.1365-2893.2001.00264.x","type":"journal-article","created":{"date-parts":[[2003,3,12]],"date-time":"2003-03-12T09:19:03Z","timestamp":1047460743000},"page":"132-138","source":"Crossref","is-referenced-by-count":3,"title":["HVR1 quasispecies analysis from a long\u2010term culture of hepatitis C virus in Hep G2 derived cells grown in a haemodialysis cartridge"],"prefix":"10.1111","volume":"8","author":[{"given":"C.","family":"Clarysse","sequence":"first","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"L.","family":"Lin","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"T.","family":"Crabb\u00e9","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"J. F.","family":"Van Pelt","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"N.","family":"Cammack","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"S. H.","family":"Yap","sequence":"additional","affiliation":[],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"311","published-online":{"date-parts":[[2003,12,15]]},"reference":[{"key":"e_1_2_6_2_2","doi-asserted-by":"publisher","DOI":"10.1056\/NEJM198911303212202"},{"key":"e_1_2_6_3_2","doi-asserted-by":"publisher","DOI":"10.1126\/science.2523562"},{"key":"e_1_2_6_4_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.87.3.983"},{"key":"e_1_2_6_5_2","doi-asserted-by":"publisher","DOI":"10.7326\/0003-4819-115-2-111"},{"key":"e_1_2_6_6_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(89)91016-7"},{"key":"e_1_2_6_7_2","first-page":"004","article-title":"Prevalence of antibodies to hepatitis C virus in Spanish patients with hepatocellular carcinoma and hepatic cirrhosis [see comments]","volume":"2","author":"Bruix J","year":"1989","journal-title":"Lancet"},{"key":"e_1_2_6_8_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.87.17.6547"},{"key":"e_1_2_6_9_2","doi-asserted-by":"publisher","DOI":"10.1002\/hep.1840140227"},{"key":"e_1_2_6_10_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.67.3.1385-1395.1993"},{"key":"e_1_2_6_11_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.87.6.2057"},{"key":"e_1_2_6_12_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.88.6.2451"},{"key":"e_1_2_6_13_2","unstructured":"HoughtonMH.Hepatitis C viruses. In: Fields BN Knipe DN & Howley PM eds.Fields Virology 3rd edn. Philadelphia: Lippincott Raven 1996: 1035\u20131058."},{"key":"e_1_2_6_14_2","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-77-5-1043"},{"key":"e_1_2_6_15_2","doi-asserted-by":"publisher","DOI":"10.1093\/infdis\/166.6.1276"},{"key":"e_1_2_6_16_2","doi-asserted-by":"publisher","DOI":"10.1016\/0168-8278(92)90073-X"},{"key":"e_1_2_6_17_2","doi-asserted-by":"publisher","DOI":"10.1182\/blood.V82.12.3701.3701"},{"key":"e_1_2_6_18_2","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-74-4-669"},{"key":"e_1_2_6_19_2","doi-asserted-by":"publisher","DOI":"10.1002\/jmv.1890420314"},{"key":"e_1_2_6_20_2","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-76-10-2485"},{"key":"e_1_2_6_21_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.89.12.5477"},{"key":"e_1_2_6_22_2","doi-asserted-by":"publisher","DOI":"10.1073\/pnas.90.13.6037"},{"key":"e_1_2_6_23_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0923-2516(06)80041-5"},{"key":"e_1_2_6_24_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.68.12.8406-8408.1994"},{"key":"e_1_2_6_25_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-8278(94)80023-5"},{"key":"e_1_2_6_26_2","doi-asserted-by":"publisher","DOI":"10.1006\/bbrc.1995.1123"},{"key":"e_1_2_6_27_2","doi-asserted-by":"publisher","DOI":"10.1128\/JVI.70.5.3325-3329.1996"},{"key":"e_1_2_6_28_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1440-1746.1995.tb01341.x"},{"key":"e_1_2_6_29_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1349-7006.1996.tb02101.x"},{"key":"e_1_2_6_30_2","doi-asserted-by":"publisher","DOI":"10.1002\/(sici)1096-9071(199602)48:2<133::aid-jmv3>3.3.co;2-#"},{"key":"e_1_2_6_31_2","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-78-10-2467"},{"key":"e_1_2_6_32_2","doi-asserted-by":"publisher","DOI":"10.1016\/s0168-1702(98)00063-x"},{"key":"e_1_2_6_33_2","doi-asserted-by":"publisher","DOI":"10.1006\/viro.1994.1381"},{"key":"e_1_2_6_34_2","first-page":"31","article-title":"Susceptibility of human liver cell cultures to hepatitis C virus infection","volume":"8","author":"Carloni G","year":"1993","journal-title":"Arch Virol Suppl"},{"key":"e_1_2_6_35_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0923-2516(06)80040-3"},{"key":"e_1_2_6_36_2","doi-asserted-by":"publisher","DOI":"10.1016\/s0923-2516(97)89901-3"},{"key":"e_1_2_6_37_2","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-79-10-2367"},{"key":"e_1_2_6_38_2","first-page":"16","article-title":"Bilirubin conjugation by an artificial liver composed of cultured cells and synthetic capillaries","volume":"21","author":"Wolf C","year":"1975","journal-title":"Trans Am Soc Artif Intern Organs"},{"key":"e_1_2_6_39_2","doi-asserted-by":"publisher","DOI":"10.1002\/hep.1840160112"},{"key":"e_1_2_6_40_2","doi-asserted-by":"publisher","DOI":"10.1111\/j.1349-7006.1997.tb00309.x"},{"key":"e_1_2_6_41_2","first-page":"151","article-title":"Immortalized human liver cell lines with phenotypical characteristics of hepatoblast and\/or biliary epithelial cells: an in vitro model for the study of liver cell\u2010specific functions","volume":"5","author":"Willems M","year":"1994","journal-title":"Transplantology"},{"key":"e_1_2_6_42_2","doi-asserted-by":"publisher","DOI":"10.1016\/s0379-0738(98)00059-0"},{"key":"e_1_2_6_43_2","doi-asserted-by":"publisher","DOI":"10.1016\/0300-2977(95)00101-8"},{"key":"e_1_2_6_44_2","doi-asserted-by":"publisher","DOI":"10.1016\/0140-6736(90)92487-3"},{"key":"e_1_2_6_45_2","doi-asserted-by":"publisher","DOI":"10.1016\/0003-2697(87)90021-2"},{"key":"e_1_2_6_46_2","doi-asserted-by":"publisher","DOI":"10.1002\/hep.1840170328"},{"key":"e_1_2_6_47_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0140-6736(94)91509-1"},{"key":"e_1_2_6_48_2","doi-asserted-by":"publisher","DOI":"10.1016\/S0168-8278(97)80347-9"},{"key":"e_1_2_6_49_2","doi-asserted-by":"publisher","DOI":"10.1099\/0022-1317-78-2-329"}],"container-title":["Journal of Viral Hepatitis"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/api.wiley.com\/onlinelibrary\/tdm\/v1\/articles\/10.1046%2Fj.1365-2893.2001.00264.x","content-type":"unspecified","content-version":"vor","intended-application":"text-mining"},{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/pdf\/10.1046\/j.1365-2893.2001.00264.x","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2023,10,17]],"date-time":"2023-10-17T13:27:38Z","timestamp":1697549258000},"score":1,"resource":{"primary":{"URL":"https:\/\/onlinelibrary.wiley.com\/doi\/10.1046\/j.1365-2893.2001.00264.x"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2001,3]]},"references-count":48,"journal-issue":{"issue":"2","published-print":{"date-parts":[[2001,3]]}},"alternative-id":["10.1046\/j.1365-2893.2001.00264.x"],"URL":"https:\/\/doi.org\/10.1046\/j.1365-2893.2001.00264.x","archive":["Portico"],"relation":{},"ISSN":["1352-0504","1365-2893"],"issn-type":[{"value":"1352-0504","type":"print"},{"value":"1365-2893","type":"electronic"}],"subject":[],"published":{"date-parts":[[2001,3]]}}}