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Titolo:
Woodchuck hepatocytes remain permissive for hepadnavirus infection and mouse liver repopulation after cryopreservation
Autore:
Dandri, M; Burda, MR; Gocht, A; Torok, E; Pollok, JM; Rogler, CE; Will, H; Petersen, J;
Indirizzi:
Univ Hamburg, Klinikum Eppendorf, Med Kernklin & Poliklin, Dept Pathol, D-20246 Hamburg, Germany Univ Hamburg Hamburg Germany D-20246 pt Pathol, D-20246 Hamburg, Germany Univ Hamburg, Klinikum Eppendorf, Dept Hepatobiliary Surg & Transplantat, D-20246 Hamburg, Germany Univ Hamburg Hamburg Germany D-20246 nsplantat, D-20246 Hamburg, Germany Univ Hamburg, Klinikum Eppendorf, Dept Med, D-20246 Hamburg, Germany Univ Hamburg Hamburg Germany D-20246 Dept Med, D-20246 Hamburg, Germany Albert Einstein Coll Med, Marion Bessin Liver Res Ctr, Bronx, NY 10467 USAAlbert Einstein Coll Med Bronx NY USA 10467 Res Ctr, Bronx, NY 10467 USA Univ Hamburg, Heinrich Pette Inst Expt Virol & Immunol, D-20246 Hamburg, Germany Univ Hamburg Hamburg Germany D-20246 & Immunol, D-20246 Hamburg, Germany
Titolo Testata:
HEPATOLOGY
fascicolo: 4, volume: 34, anno: 2001,
parte:, 1
pagine: 824 - 833
SICI:
0270-9139(200110)34:4<824:WHRPFH>2.0.ZU;2-A
Fonte:
ISI
Lingua:
ENG
Soggetto:
HEPATITIS-B-VIRUS; PRIMARY RAT HEPATOCYTES; CLOSED CIRCULAR DNA; IN-VITRO; IMMUNOSUPPRESSED PATIENTS; FUNCTIONAL-ANALYSIS; PRIMARY CULTURE; DMSO CULTURE; X PROTEIN; TRANSPLANTATION;
Tipo documento:
Article
Natura:
Periodico
Settore Disciplinare:
Clinical Medicine
Life Sciences
Citazioni:
41
Recensione:
Indirizzi per estratti:
Indirizzo: Petersen, J Univ Hamburg, Klinikum Eppendorf, Med Kernklin & Poliklin, Dept Pathol, Martinistr 52, D-20246 Hamburg, Germany Univ Hamburg Martinistr 52 Hamburg Germany D-20246 g, Germany
Citazione:
M. Dandri et al., "Woodchuck hepatocytes remain permissive for hepadnavirus infection and mouse liver repopulation after cryopreservation", HEPATOLOGY, 34(4), 2001, pp. 824-833

Abstract

Isolated hepatocytes represent a relevant model of the liver and are highly required both for research and therapeutic applications. However, sourcesof primary liver cells from human beings and from some animal species are limited. Therefore, cryopreservation of hepatocytes could greatly facilitate advances in various research areas. The aim of this study Was to evaluatewhether cryopreserved primary woodchuck hepatocytes could be used for woodchuck hepatitis B virus, (WHV) infection studies, and whether they could maintain their regenerative potential in vivo after thawing. Critical steps for good quality of cryopreserved hepatocytes included the use of Universityof Wisconsin (UW) solution as a main component of the freezing medium, stepwise reduction of dimethylsulfoxide (DMSO) to avoid osmotic shock, and maintenance of low concentrations of DMSO in the culture medium. After cryopreservation, cell viability was still high (70% to 80%), and 50% to 60% of thawed cells attached to the plates. The appearance of covalently closed circular (ccc)DNA and of WHV-replicative forms a few days after in vitro infection demonstrated that thawed woodchuck hepatocytes were still susceptible to viral infection, thus proving maintenance of a very high hepatocyte-specific differentiation status. Furthermore, transplantation of woodchuck hepatocytes into the liver of urokinase-type plasminogen activator (uPA)/recombination activation gene-2 (RAG-2) mice, a model of liver regeneration, demonstrated that cryopreserved cells retained the ability to divide and to extensively repopulate a xenogenic liver. Notably, in vivo susceptibility to infection with WHV and proliferative capacity of frozen/thawed woodchuck hepatocytes in recipient mice were identical to those observed by transplantingfresh hepatocytes.

ASDD Area Sistemi Dipartimentali e Documentali, Università di Bologna, Catalogo delle riviste ed altri periodici
Documento generato il 23/09/20 alle ore 12:11:13