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Creating an In Vivo System to Study Cell Cycle Regulation of Pluripotent Stem Cell Differentiation During Mammalian Gastrulation Aaron Cohen, Rebecca Kim-Yip, Eszter Posfai Gastrulation is a significant morphogenetic event in early mammalian development when precursor pluripotent epiblast cells migrate through the primitive streak and differentiate into mesoderm, endoderm, and primordial germ cells. The formation of these distinct cell types is required for the proper development of the organism's tissues, organs, and germline. Not only do epiblast cells migrate and differentiate during gastrulation, but they also rapidly proliferate. This suggests that careful coordination of these processes must exist to ensure the embryo develops with the proper cell number and types. Recent evidence in human pluripotent stem cells (PSC) suggests that cell cycle progression interacts with cell fate specification during gastrulation. However, a significant limitation of these studies has been the inability to create an aligned in vivo model. To investigate the cell cycle's role in coordinating differentiation, I will use PIP-FUCCI, a well characterized live reporter of cell cycle phase. I will study the cell cycle's role in coordinating germ layer and primordial germ cell specification in vitro using mouse embryonic stem cells. Then, I will create a reliable in vivo system via a transgenic mouse line to validate findings in a gastrulating mouse embryo. This summer, I have made progress on establishing a formative state PSC line that I will use to investigate cell cycle regulation of PSC differentiation during gastrulation. I have also made progress on characterizing and validating an in vivo reporter of PSC differentiation. My findings will expand our understanding of the mechanisms of cell cycle coordination of cell differentiation and germ layer formation during gastrulation and mammalian development.

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Princeton

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