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dc.contributor.authorMurray, Erica
dc.date.accessioned2011-08-18 17:04:49 (GMT)
dc.date.available2011-08-18 17:04:49 (GMT)
dc.date.issued2011-08-18T17:04:49Z
dc.date.submitted2011
dc.identifier.urihttp://hdl.handle.net/10012/6096
dc.description.abstractRegenerative medicine is a rapidly developing field, merging engineering and biological life sciences to create biological replacements for damaged tissue and organ function. Development of cellular based therapies has the potential of curing present untreatable diseases and conditions, such as diabetes. The identification of protein expression patterns, that guide undifferentiated cells to different lineages, can provide important information about the progression of cellular differentiation at various stages. This research project utilizes proteomics and in vitro live-cell microscopy to investigate two distinct cellular systems: (1) the signaling pathways of calmodulin (CaM) in the differentiation of a human glioblastoma cell line; and (2) the effect of islet neogenesis associated protein (INGAP) on human islet-derived progenitor cells (hIPCs). Using a proteomic readout with a long term live-cell imagining approach, it was hypothesized that highly specific binding proteins of a CaM-mutant, and proteins in hIPCs perturbed by INGAP, could be identified and studied in vitro, characterizing specific signaling pathways which control the function of CaM in brain tumour cells and the mechanism(s) of INGAP in islet-derived progenitor cells. This thesis presents the utility of a proteomics and an in vitro cell microscopy approach to investigate therapeutic proteins, such as INGAP, on cell culture systems. The results have established the limitations and the utility of DIGE, differential binding of a CaM-mutant versus calcium-CaM, and the cell specific uptake feasibility of using the TAT-binding domain. In the hIPC system, proteomic, phenotypic, motility, proliferation and nuclear effects of INGAP were determined. Specifically, hIPCs exposed to INGAP had 50% decrease in average nuclear speed, the translocation of two identified proteins caldesmon and tropomyosin and INGAP was found to bind specifically to hIPCs. However, hIPCs had no changes in insulin specific hormone expression.en
dc.language.isoenen
dc.publisherUniversity of Waterlooen
dc.subjectcalmodulinen
dc.subjecthuman islet derived progenitor cellsen
dc.subjectdifferential in gel electrophoresisen
dc.subjectlive-cell imagingen
dc.subjectstem cellen
dc.subjectdifferentiationen
dc.subjectproteomicsen
dc.subjectcaldesmonen
dc.subjecttropomyosinen
dc.titleProteomic Analysis and Long Term Live Cell Imaging of Primary Human Cells in Cultureen
dc.typeDoctoral Thesisen
dc.pendingfalseen
dc.subject.programChemical Engineeringen
uws-etd.degree.departmentChemical Engineeringen
uws-etd.degreeDoctor of Philosophyen
uws.typeOfResourceTexten
uws.peerReviewStatusUnrevieweden
uws.scholarLevelGraduateen


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