School of Biomedical Sciences, The University of Queensland, Brisbane, QLD, Australia.
Institute for Molecular Bioscience, The University of Queensland, Brisbane, QLD, Australia.
Methods Mol Biol. 2023;2677:221-231. doi: 10.1007/978-1-0716-3259-8_13.
The fetal gonad contains a great variety of differentiating cell populations, of which germ cells make up a relatively small percentage. In order to study germ cell-specific gene and protein expression, as well as determine direct effects of signaling molecules, it is necessary to prepare enriched populations of germ cells and maintain them in culture for several hours to multiple days. The protocols in this chapter are designed to provide a guide for the isolation or enrichment of primordial germ cells (from 9.5 days post coitum (dpc) to 18.5 dpc) by flow cytometry (Subheading 3.1) or magnetic sorting (Subheading 3.2), followed by feeder-free primary germ cell culture (Subheading 3.3).
胎儿生殖腺包含多种分化细胞群体，其中生殖细胞占相对较小的比例。为了研究生殖细胞特异性基因和蛋白质表达，并确定信号分子的直接影响，有必要通过流式细胞术（见 3.1 小节）或磁分选（见 3.2 小节）来分离或富集原始生殖细胞（从 9.5 天妊娠龄（dpc）到 18.5 dpc），并在无饲养层的原代生殖细胞培养（见 3.3 小节）中对其进行培养。本章的方案旨在提供通过流式细胞术（小节 3.1）或磁分选（小节 3.2）分离或富集原始生殖细胞（从 9.5 天妊娠龄（dpc）到 18.5 dpc）的指南，然后进行无饲养层的原代生殖细胞培养（小节 3.3）。
