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.
Elife. 2022 Apr 8;11:e72811. doi: 10.7554/eLife.72811.
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.
人类原始生殖细胞(hPGCs)大约在着床时形成,是卵子和精子的前体细胞。由于早期着床后人类胚胎无法用于研究,因此 hPGC 特化的许多方面仍未得到很好的理解。在这里,我们表明,用 BMP4 处理的微图案化人类多能干细胞(hPSC)会产生 hPGC 样细胞(hPGCLC),并将其用作一种定量重现且简单的体外模型来探究这一重要的发育事件。我们对微图案化 hPSC 进行了长达 96 小时的研究,并表明 hPGCLC 群体是稳定的,并继续成熟。通过在 hPGCLC 分化过程中干扰信号,我们确定了 Nodal 信号的一个以前未被认识到的作用,并发现 BMP 和 Nodal 信号的相对时间和持续时间是控制 hPGCLC 数量的关键参数。我们提出了一个由 Nodal 和 BMP 信号驱动的交叉抑制命运的网络数学模型,该模型预测了信号干扰后的测量命运模式。最后,我们表明 hPSC 集落大小决定了 hPGCLC 特化的效率,这促使我们大大提高了 hPGCLC 分化的效率。