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|Title: ||The Epigenetics of Pluripotency in Embryonic Stem Cells|
|Authors: ||Antoninus, Andreas Ardhika|
|Issue Date: ||31-Aug-2015|
|Publisher: ||University of Sydney|
Sydney Medical School
|Abstract: ||The utilisation of pluripotent cells in regenerative medicine requires the cells to be faithfully maintained in a pluripotent state. One epigenetic feature associated with pluripotency in the embryo is the relative global hypomethylation of cytosine (at CpG dinucleotides, 5meC) that occurs within the nucleus of the inner cell mass and epiblast. It is surprising therefore that the global levels of 5meC in embryonic stem cells (ESCs) propagated under conventional methods are as high as many differentiated somatic cells. Using the newly developed antigen retrieval method that incorporates both acid and trypsin based epitope retrieval processes this project assessed the effects of culture conditions on global levels and localization of 5meC in ESCs. The study shows that the loss of global DNA methylation accompanied increased pluripotency achieved by ESC in 2i media or in suspension culture of EBs. It shows for the first time that this was accompanied by the recruitment of 5meC to heterochromatin. It also showed that global hypermethylation is an early response to conditions that favour the loss of pluripotency, and occurs before evidence of the loss pluripotency or morphological differentiation. This finding provides a new framework for further investigation of the epigenetic requirements of the pluripotent state.|
|Type of Work: ||Masters Thesis|
|Type of Publication: ||Master of Philosophy M.Phil|
|Appears in Collections:||Sydney Digital Theses (Open Access)|
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|ANTONINUS Andreas - Final Thesis.pdf||Thesis||7.01 MB||Adobe PDF|
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