Type of Document	Master's Thesis
Author	Coley, Laura Whitney
Author's Email Address	lcoley1@tigers.lsu.edu
URN	etd-11182010-204302
Title	Cell Extract-Based Reprogramming of Somatic Cells
Degree	Master of Science (M.S.)
Department	Animal Science (Animal, Dairy, & Poultry Sciences)
Advisory Committee	Advisor Name Title Bondioli, Kenneth Committee Chair Dimario, Patrick Committee Member Eilersten, Kenneth Committee Member Godke, Robert Committee Member
Keywords	Cell extracts
Date of Defense	2010-11-05
Availability	unrestricted
Abstract The differentiation potential of adult stem cells (ASC) has long been thought to be limited to cell lineages present in the organ from which they are derived; however, several studies have challenged this notion by demonstrating that some ASC exhibit a remarkably high degree of plasticity. Unlike terminally differentiated somatic cells, the less differentiated state of ASC can assume the functional phenotypes and expression profiles of cells unique to other tissues. The expansive repertoire of differentiation potential exhibited by ASC suggests these cells possess characteristics similar to pluripotent cells, including epigenomic regulatory pattern. Therefore, ASC may be better equipped for complete epigenetic reprogramming than terminally differentiated cells. The objective of Experiment 1 was to analyze bovine adipose-derived adult stem cells (ADAS) and fetal fibroblast (BFF) cells for the presence of the pluripotency-associated genes, Oct-4, Nanog, and Sox-2. Because the endogenous expression of these genes is believed to contribute to reprogramming efficiency, Experiment 2 sought to increase Oct-4, Nanog and Sox-2 expression levels in BFF cells through exposure to ADAS cell extracts. Transcripts for all three pluripotency-associated genes were detected in all BFF and ADAS cell samples at every passage analyzed; however, expression was quite low and highly variable between cell lines and passage numbers. Nevertheless, these findings support the notion that these cells are less differentiated than other somatic cells. This less differentiated state appears to sufficient for at least the partial reprogramming of BFF cells using ADAS cell extracts in a cell extract-based nuclear reprogramming system.
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