(261f) Exploring the Role of Energy Metabolism In the Differentiation of Pluripotent Stem Cells towards Hepatic Fate

Hepatocytes, the main parenchymal cell type of the liver, perform several important functions including detoxification and maintenance of glucose homeostasis via processes such as glycogenesis and gluconeogenesis. Stem cell derived hepatocytes hold great potential for applications in the development of pharmaceuticals and cell therapy. We have previously demonstrated that pluripotent stem cells can be guided towards a hepatic fate, yielding cells that exhibit key hepatocyte functions. In this study, specific promoter-reporter lines were generated for monitoring the progress of differentiation as well as enriching desired cell populations, including both progenitor and mature hepatocyte-like cells, by cell sorting. Gene expression data of the enriched cells was utilized to elucidate the key players involved in hepatic differentiation and maturation. The cells express key hepatic transcripts including drug metabolizing CYP450s and exhibit functional capabilities such as albumin production and urea secretion. Furthermore, this transition is likely to be accompanied by changes in metabolism and as expected, the enriched hepatocyte like-cells begin to demonstrate a switch towards the expression of key enzyme isoforms, partaking in liver-specific functions. Given the isoform specific allosteric regulation of the metabolic enzymes and the extensive crosstalk that occurs between metabolic and signaling networks, this system allows for probing the relationship between metabolism and differentiation. These findings provide insights into the complex liver development process and allow for optimization of conditions to enhance differentiation potential and maturity.