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Differentiation of mouse embryonic stem cells along a hepatocyte lineage
Descriptive
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Title
Differentiation of mouse embryonic stem cells along a hepatocyte lineage
Name
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Novik
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Eric I.
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Eric I. Novik
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author
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Yarmush
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Martin
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Advisory Committee
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Martin l Yarmush
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chair
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Schloss
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Rene
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Advisory Committee
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Rene Schloss
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internal member
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Roth
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Charles
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Advisory Committee
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Charles Roth
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internal member
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Tilles
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Arno
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Advisory Committee
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Arno Tilles
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outside member
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Rutgers University
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degree grantor
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Graduate School - New Brunswick
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theses
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2007
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2007-10
Language
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English
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electronic
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x, 67 pages
Abstract
The development of implantable engineered liver tissue constructs, ex vivo hepatocyte based therapeutic devices and drug discovery studies are limited by an inadequate hepatocyte cell source. Embryonic stem (ES) cells, characterized by their self-renewing and multi-lineage differentiating capabilities, represent a promising mature cell source required for these applications. Previous research has utilized embryoid body (EB) formation in both guided, through extracellular matrix and growth factor supplementation, and unguided, or spontaneous, differentiation to generate hepatocyte like cells. However, these characterizations have been limited to only one or several lineage specific protein or gene expression patterns. In addition, there have been few reports of long term propagation or characterization of long term function for ES cell derived hepatocyte precursors. In this thesis, we have implemented a platform for the long term propagation and augmentation of functional hepatocytes generated from murine ES cell sources. We first utilize a controlled, reproducible, EB mediated differentiation system to characterize efficiency of hepatocyte lineage commitment in four parallel culture configurations. These studies have shown that, EB mediated stem cell differentiation spontaneously yield populations of hepatocyte lineage cells expressing mature hepatocyte markers such as albumin (ALB) and Cytokeratin 18 (CK-18). We then used secondary culture configurations to study the effects of collagen sandwich culture and Oncostatin-M (OSM) or S-nitroso-N-acetylpenicillamine (SNAP) supplementation of EB derived hepatocyte-lineage cell function. The results of these studies suggest that SNAP, independent of the collagen supplementation, maintains the highest levels of ALB expression, however mature liver specific CK-18 is only expressed in the presence of both gel sandwich culture supplemented with SNAP. In addition, albumin secretion and Cytochrome P450 detoxification studies indicated that this condition was the best for the augmentation of hepatocyte-like function. Maintenance and augmentation of hepatocyte-like cells isolated from heterogeneous EB cell populations will be a critical step in generating large numbers of functional differentiated cells for therapeutic use.
Note
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Ph.D.
Note
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Includes bibliographical references (p. 45-48).
Subject
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Topic
Biomedical Engineering
Subject
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Topic
Liver cells--Differentiation
Subject
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(authority = ETD-LCSH)
Topic
Embryonic stem cells--Differentiation
Subject
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(authority = ETD-LCSH)
Topic
Cell differentiation
Subject
(ID = SUBJ5);
(authority = ETD-LCSH)
Topic
Stem cells
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Title
Graduate School - New Brunswick Electronic Theses and Dissertations
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rucore19991600001
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http://hdl.rutgers.edu/1782.2/rucore10001600001.ETD.17086
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ETD_275
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Identifier
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doi:10.7282/T3NG4R0F
Genre
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ETD doctoral
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Rights
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The author owns the copyright to this work.
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Copyright protected
Availability
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Open
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Name
Eric Novik
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Copyright holder
Affiliation
Rutgers University. Graduate School - New Brunswick
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Non-exclusive ETD license
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Author Agreement License
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I hereby grant to the Rutgers University Libraries and to my school the non-exclusive right to archive, reproduce and distribute my thesis or dissertation, in whole or in part, and/or my abstract, in whole or in part, in and from an electronic format, subject to the release date subsequently stipulated in this submittal form and approved by my school. I represent and stipulate that the thesis or dissertation and its abstract are my original work, that they do not infringe or violate any rights of others, and that I make these grants as the sole owner of the rights to my thesis or dissertation and its abstract. I represent that I have obtained written permissions, when necessary, from the owner(s) of each third party copyrighted matter to be included in my thesis or dissertation and will supply copies of such upon request by my school. I acknowledge that RU ETD and my school will not distribute my thesis or dissertation or its abstract if, in their reasonable judgment, they believe all such rights have not been secured. I acknowledge that I retain ownership rights to the copyright of my work. I also retain the right to use all or part of this thesis or dissertation in future works, such as articles or books.
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