Department of Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, United States; Department of Biomedical Engineering, University of Michigan, Ann Arbor, United States; Center for Organogenesis, University of Michigan Medical School, Ann Arbor, United States; Department of Internal Medicine, Gastroenterology, University of Michigan Medical School, Ann Arbor, United States
Department of Cell and Developmental Biology, University of Michigan Medical School, Ann Arbor, United States; Department of Biomedical Engineering, University of Michigan, Ann Arbor, United States; Center for Organogenesis, University of Michigan Medical School, Ann Arbor, United States; Department of Physics, University of Michigan, Ann Arbor, United States
Human primordial germ cells (hPGCs) form around the time of implantation and are the precursors of eggs and sperm. Many aspects of hPGC specification remain poorly understood because of the inaccessibility of the early postimplantation human embryo for study. Here, we show that micropatterned human pluripotent stem cells (hPSCs) treated with BMP4 give rise to hPGC-like cells (hPGCLC) and use these as a quantitatively reproducible and simple in vitro model to interrogate this important developmental event. We characterize micropatterned hPSCs up to 96 hr and show that hPGCLC populations are stable and continue to mature. By perturbing signaling during hPGCLC differentiation, we identify a previously unappreciated role for Nodal signaling and find that the relative timing and duration of BMP and Nodal signaling are critical parameters controlling the number of hPGCLCs. We formulate a mathematical model for a network of cross-repressive fates driven by Nodal and BMP signaling, which predicts the measured fate patterns after signaling perturbations. Finally, we show that hPSC colony size dictates the efficiency of hPGCLC specification, which led us to dramatically improve the efficiency of hPGCLC differentiation.
