Wellcome Trust/Cancer Research UK Gurdon Institute, University of Cambridge, Cambridge, UK.
Department of Physiology, Development and Neuroscience, University of Cambridge, Cambridge, UK.
Methods Mol Biol. 2021;2214:75-89. doi: 10.1007/978-1-0716-0958-3_6.
The specification and development of germ cells to gametes is a unique process, which is of great biological and clinical relevance. In mammals, the founding cells of the germline are primordial germ cells (PGCs), which arise during early embryogenesis. The low number of PGCs within the developing embryo limits the study of these cells in model organisms. The generation of PGC-like cells (PGCLCs) from murine pluripotent stem cells reconstitutes the earliest stages of germ cell development and mitigates the technical constraints of studying this developmental process in vivo. Here, we describe the technical details of the PGCLC specification approach and illustrate adaptations designed to improve compatibility with methods such as chromatin immunoprecipitation by increasing the yield of PGCLC generation.
生殖细胞到配子的特化和发育是一个独特的过程，具有重要的生物学和临床意义。在哺乳动物中，生殖系的创始细胞是原始生殖细胞（PGCs），它们在胚胎早期出现。发育中的胚胎中 PGC 的数量较少，限制了在模式生物中对这些细胞的研究。从鼠多能干细胞中生成 PGC 样细胞（PGCLC）可重建生殖细胞发育的最早阶段，并减轻了在体内研究该发育过程的技术限制。在这里，我们描述了 PGCLC 特化方法的技术细节，并说明了为提高与染色质免疫沉淀等方法的兼容性而设计的适应性改进，以增加 PGCLC 生成的产量。
