Date of Award
Master of Science (MS)
Molecular Science and Nanotechnology
Human adipose-derived mesenchymal stem cells offer significant therapeutic potential as an ethically sound, easily accessed source of adult stem cells. To harness their medically relevant properties, it is necessary to understand the mechanisms that control their fate. Cell state and differentiation of stem cells is determined by interactions of signaling pathways, chromatin modifiers, and transcription factors working to regulate cell-type specific gene expression profiles. Specifically, both the MED12 subunit of the Mediator complex and the Notch signaling pathway are known to individually influence hASC self-renewal. We investigated the relationship between Notch signaling and transcriptional cofactor, MED12, to elucidate a potential regulatory relationship and better understand the mechanisms that determine cell fate in hASCs.
Using siRNA mediated knockdowns, we analyzed the expression and activation changes of Notch signaling in self-renewing adipose stem cells in the presence of reduced MED12. Knockdown validation, Notch expression and signaling pathway activation was quantitated via qRT-PCR and western blot analysis. Subsequently, we observed that MED12 is required for the activation of the Notch3 signaling pathway, while Notch1 signaling is not significantly influenced by the reduction of MED12, suggesting a novel regulatory interface between MED12 and the Notch3 signaling pathway. Understanding the relationship between MED12, Notch1, and Notch3 and their influence on self-renewal will increase understanding of hASC cell fate mechanisms, indicating a need for further investigation, and aiding in better determining their potential for applications in regenerative medicine.
Furthermore, to advance support for scientific investigation and essential stem cell research, public education of the basic science and medical relevance of stem cells must also be addressed. An interactive children’s book was developed to integrate basic science research and stem cell concepts inside and out of formal educational facilities. Specifically designed as an educational book for students, a tool for educators, and a resource for the community, this book aims to effectively communicate fact-based stem cell content, address misconceptions, and promote positive engagement and interest in the sciences.
This thesis provides evidence of a novel regulatory relationship between Notch signaling and MED12 and contributes, via educational resource, to advancing support for essential stem cell research. Collectively, this aids in the elucidating the potential use and benefits of adipose stem cells in clinical therapeutics and regenerative medicine.
Hodnett, Rebecca, "" (2021). Thesis. 49.