Date of Award
Master of Science (MS)
Induced pluripotent stem cells (iPSCs) can be differentiated into any cell type found in the body. The derivation of a stem cell derived β cell (SC-β) capable of responding to glucose by secreting insulin was hugely significant for diabetes research and opened up the possibility of cell replacement therapy to combat this widespread disease (Pagliuca et al. 2014). The optimization of differentiation procedures such as this could improve yield, function, cost, and efficiency of a stem cell-derived product. Current approaches to improve differentiation are primarily focused on signal transduction pathways, while the metabolic state of the cells has received little attention. Using a Seahorse XF24 extracellular flux analyzer, we found that iPSCs have heavy reliance on fatty acid oxidation (FAO), a trait not shared by many mature cell types. Manipulation of stem cells and their mitochondrial oxidation by manipulating cell culture composition or adding small molecules BPTES, etomoxir, and UK5099 was shown to be possible in both suspension and adherent differentiations. Expression of key mitochondrial genes changed drastically during differentiation of iPSCs to SC-β cells. Thus, this body of work has demonstrated the importance of and paved the way for future research on the importance of mitochondrial oxidation to differentiation.
Jeffrey Millman, Irfan Lodhi, Dennis Barbour