Investigation of transcriptional control in directing cell fate

University Hall Lobby

Cell fate is driven by changes in gene expression which are the result of external cues, endogenous signaling pathways, and transcriptional machinery all working together. Human adipose-derived stem cells (hASCs) are adult stem cells derived from adipose tissue that are used in clinical research because they can be easily extracted, are multipotent, are able to self-renew, and have natural immunomodulatory capabilities. Culturing hASCs in the lab allows for the in-depth study of cell fate determination with the ability to manipulate the environment and easily assay changes in cell fate, transcription, and protein expression. Our project is primarily interested in investigating how protein expression levels of the four subunits of the Mediator kinase domain affect adipogenesis of hASCs. We are using cell culture to grow and examine changes in cell behavior over seven days during which cells are stimulated to undergo adipogenesis. Gene expression is assessed through the collection of RNA which is then used to synthesize cDNA, that is analyzed using gene specific primers to measure relative transcript levels using both end-point and quantitative RT-PCR. Protein expression is quantified and analyzed through a Bradford Assay and Western Blot using specific antibodies to assess Mediator protein levels. By using cell culture, directed differentiation, qRT-PCR, and western blots we are able to understand the transcriptional processes that directs hASC fate, creating a strong foundation for future research to investigate novel therapies for cancers and other disorders.