Date of Award
Master of Science
Dr. Gilles Lajoie
Dr. Lynne Postovit
Hypoxia has been shown to promote pluripotency in human embryonic stem cells (hESCs), but the mechanism by which this occurs in poorly understood. To gain insight into this mechanism, we used mass spectrometry to investigate changes in protein expression in hESCs cultured in hypoxia. hESCs in feeder free culture were incubated in 1% oxygen or 20% oxygen for 48 and 72 hours. The medium was not changed during this time to accelerate differentiation. Immunofluroescence localization of Oct-4 revealed that cultures incubated in hypoxia were less differentiated than cultures incubated in normoxia. Electrospray tandem mass spectrometry was performed to compare global protein expression of hESCs from each oxygen condition. Changes were observed in the expression of proteins involved in metabolism, chromatin modification, post-transcriptional modification, and regulation of the transcription factor, c-Myc. The results of this study will improve our understanding of the mechanism by which hypoxia maintains pluripotency of hESCs in vitro.
Nuhn, Amelia A., "Proteomic studies of human embryonic stem cells in hypoxia" (2011). Digitized Theses. 3475.