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Use of gene expression to characterize heterogeneous liver cell populations

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dc.contributor.advisor Linda Griffith. en_US Schreiber, Brent M. (Brent Matthew), 1981- en_US
dc.contributor.other Massachusetts Institute of Technology. Biological Engineering Division. en_US 2005-06-02T19:45:25Z 2005-06-02T19:45:25Z 2004 en_US
dc.description Thesis (M. Eng.)--Massachusetts Institute of Technology, Biological Engineering Division, 2004. en_US
dc.description Includes bibliographical references (leaves 84-93). en_US
dc.description.abstract Non-parenchymal cells (NPC's) are integral to recreate the native hepatic microenvironment and necessary to maintain in vivo liver function. A variety of in vitro culture systems have been developed to address different aspects of liver physiology and architecture in order to recreate the microenvironment. These in vitro co-culture strategies have been limited by their inability to systematically characterize the addition of non-parenchymal cells. In this dissertation, I use gene expression levels quantified by real-time RT-PCR to determine tissue composition. The identified genes demonstrate significant cell-type specificity, magnitude, and stability of expression in vivo and throughout each step of cell isolation process. In the course of this development, we establish protocols to accurately isolate and count an enriched fraction of primary NPC's. Experiments on the perfusion and isolation process prove that there exists an inverse correlation between perfusion flow rate and NPC yield and viability. Further, we have characterized the tissue composition of each step in the cell isolation process and the resulting NPC population to confirm that a significant number of each NPC type is delivered to in vitro co-culture. System output analysis of spheroids co-cultured at physiological ratios and seeded into the milliF bioreactor shows the presence of stellate cells, but the absence of endothelial (EC) and kupffer cells (KC). The same analysis of 2D collagen gel sandwiches shows the presence of all NPC cell types. This indicates that our process is currently limited by the ability of EC's and KC's to incorporate into spheroid aggregates. Future work that validates the temporal expression stability of the identified genes in different in vitro culture systems en_US
dc.description.abstract (cont.) and environments will enable determination of relative levels of NPC incorporation and will allow correlations to be made between operational features of in vitro systems, the resulting culture microenvironment, and observed tissue function. en_US
dc.description.statementofresponsibility by Brent M. Schreiber. en_US
dc.format.extent 120 leaves en_US
dc.format.extent 8006001 bytes
dc.format.extent 8020925 bytes
dc.format.mimetype application/pdf
dc.format.mimetype application/pdf
dc.language.iso eng en_US
dc.publisher Massachusetts Institute of Technology en_US
dc.rights MIT theses are protected by copyright. They may be viewed, downloaded, or printed from this source but further reproduction or distribution in any format is prohibited without written permission. en_US
dc.subject Biological Engineering Division. en_US
dc.title Use of gene expression to characterize heterogeneous liver cell populations en_US
dc.title.alternative Characterization of heterogeneous hepatic cell population via use of gene expression en_US
dc.type Thesis en_US M.Eng. en_US
dc.contributor.department Massachusetts Institute of Technology. Biological Engineering Division. en_US
dc.identifier.oclc 57364170 en_US

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