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Immortalized human hepatocyte, HCV, SCID-uPA mice, de-immortalization, tamoxifen, Cre recombinase, JFH1, viral entry, HCV life cycle

Mohajerani, Seyed Amir

Supervisor and department: Mason,AndrewMedicine Douglas,DonnaSurgery Kneteman,Norman M Surgery

Examining committee member and department: Kneteman,Norman M Surgery Mason,AndrewMedicine Douglas,DonnaSurgery

Department: Department of Surgery

Specialization:

Date accepted: 2010-12-02T18:38:19Z

Graduation date: 2011-06

Degree: Master of Science

Degree level: Master's

Abstract: The chimeric Alb-uPA SCID mouse that has been transplanted with human hepatocytes is a model to facilitate in vivo study of HCV. We explored further development of the model by using repopulation with immortalized human hepatocytes IHH in place of primary human hepatocyte PHH transplantation to support HCV infection. In vitro HCV studies typically utilize a human hepatoma cell line Huh7 and rely on transfection with transcribed genomic RNA derived from a unique HCV strain JFH1. Unfortunately, this system has not been successful in support of infection with serum-derived HCV HCVser. IHH may offer an alternative since their differentiation status remains close to that of PHH. IHH transfected with HCV RNA H77 or JFH1 or infected with HCVser showed stable intracellular and supernatant HCV RNA by real-time RT-PCR. IHH showed intracellular HCV NS3 proteins. HCV transfected or infected IHH secrete infectious HCVcc for in vivo and vitro.

Language: English

DOI: doi:10.7939-R3JQ6B

Rights: License granted by Seyed Amir Mohajerani sm44@ualberta.ca on 2010-11-27T17:40:51Z GMT: Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only. Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of the above terms. The author reserves all other publication and other rights in association with the copyright in the thesis, and except as herein provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the author's prior written permission.





Autor: Mohajerani, Seyed Amir

Fuente: https://era.library.ualberta.ca/


Introducción



University of Alberta Immortalized Human Hepatocytes, an Alternate Model for the Study of the Propagation of HCV in Vivo and in Vitro By Seyed Amir Mohajerani A thesis submitted to the Faculty of Graduate Studies and Research in partial fulfillment of the requirements for the degree of Master of Science Experimental Surgery Department of Surgery ©Seyed Amir Mohajerani Spring 2011 Edmonton, Alberta Permission is hereby granted to the University of Alberta Libraries to reproduce single copies of this thesis and to lend or sell such copies for private, scholarly or scientific research purposes only.
Where the thesis is converted to, or otherwise made available in digital form, the University of Alberta will advise potential users of the thesis of these terms. The author reserves all other publication and other rights in association with the copyright in the thesis and, except as herein before provided, neither the thesis nor any substantial portion thereof may be printed or otherwise reproduced in any material form whatsoever without the authors prior written permission. Committee Members Dr.
Norman Kneteman, Surgery Dr.
Donna Douglas, Surgery Dr.
Andrew Mason, Medicine Dr.
Thomas A Churchill, Surgery Dedicated to My wife, parents and family members for their support ABSTRACT The chimeric Alb-uPA SCID mouse that has been transplanted with human hepatocytes is a model to facilitate in vivo study of HCV.
We explored further development of the model by using repopulation with immortalized human hepatocytes (IHH) in place of primary human hepatocyte (PHH) transplantation to support HCV infection.
In vitro HCV studies typically utilize a human hepatoma cell line (Huh7) and rely on transfection with transcribed genomic RNA derived from a unique HCV strain (JFH1).
Unfortunately, this system has not been successful in support of infection with serum-derived HCV (HCVser).
IHH may offer an alternative since their differentiation status remains close to that of P...





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