Master of Science
Human pluripotent stem cells (hPSCs) exhibit two unique characteristics: pluripotency and self-renewal. These properties are maintained by a series of complex signaling pathways, however, quantitative data for the respective proteins is lacking. Selected reaction monitoring (SRM) is a targeted, quantitative technique in mass spectrometry that is highly sensitive in peptide detection. In this thesis, an SRM protocol was developed in order to detect and quantify a defined set of proteins responsible for maintaining stem cell pluripotency. Two hESC differentiation protocols were validated for use as model systems within which to measure differential protein expression by SRM. SRM assays were generated for thirty-three proteins and tested on cell lysates. Wnt1 and β-catenin were shown to be upregulated during differentiation, while other proteins and peptides were detected but not quantifiable. The results of this study highlight the complexity of hPSC proteome and help further the understanding of the mechanisms responsible for pluripotency.
Kania, Kevin Gregory, "Targeted Proteomics of Human Pluripotent Stem Cells" (2014). Electronic Thesis and Dissertation Repository. 1990.