Key Laboratory of Animal Breeding Reproduction and Molecular Design for Jiangsu Province, College of Animal Science and Technology, Yangzhou University, Yangzhou 225009, China.
Institutes of Agricultural Science and Technology Development, Yangzhou University, Yangzhou 225009, China.
Genes (Basel). 2023 Oct 22;14(10):1975. doi: 10.3390/genes14101975.
Primordial germ cells (PGCs) are essential for the genetic modification, resource conservation, and recovery of endangered breeds in chickens and need to remain viable and proliferative in vitro. Therefore, there is an urgent need to elucidate the functions of the influencing factors and their regulatory mechanisms. In this study, PGCs collected from Rugao yellow chicken embryonic eggs at Day 5.5 were cultured in media containing 0, 5, 10, 20, 50, and 100 μg/mL insulin. The results showed that insulin regulates cell proliferation in PGCs in a dose-dependent way, with an optimal dose of 10 μg/mL. Insulin mediates the mRNA expression of cell cycle-, apoptosis-, and ferroptosis-related genes. Insulin at 50 μg/mL and 100 μg/mL slowed down the proliferation with elevated ion content and GSH/oxidized glutathione (GSSG) in PGCs compared to 10 μg/mL. In addition, insulin activates the PI3K/AKT/mTOR pathway dose dependently. Collectively, this study demonstrates that insulin reduces apoptosis and ferroptosis and enhances cell proliferation in a dose-dependent manner via the PI3K-AKT-mTOR signaling pathway in PGCs, providing a new addition to the theory of the regulatory role of the growth and proliferation of PGC in vitro cultures.
原始生殖细胞（PGCs）对于鸡的遗传修饰、资源保护和濒危品种的恢复至关重要，需要在体外保持存活和增殖。因此，迫切需要阐明影响因素的功能及其调节机制。在这项研究中，从 Rugao 黄鸡胚胎蛋第 5.5 天收集的 PGCs 在含有 0、5、10、20、50 和 100μg/mL 胰岛素的培养基中进行培养。结果表明，胰岛素以剂量依赖的方式调节 PGCs 的细胞增殖，最佳剂量为 10μg/mL。胰岛素介导细胞周期、凋亡和铁死亡相关基因的 mRNA 表达。与 10μg/mL 相比，50μg/mL 和 100μg/mL 的胰岛素会减缓增殖速度，同时增加 PGCs 中的离子含量和 GSH/氧化型谷胱甘肽（GSSG）。此外，胰岛素还会依赖剂量激活 PI3K/AKT/mTOR 通路。综上所述，本研究表明，胰岛素通过 PI3K-AKT-mTOR 信号通路以剂量依赖的方式减少 PGCs 中的细胞凋亡和铁死亡，并增强细胞增殖，为 PGC 在体外培养中的生长和增殖的调节作用理论提供了新的补充。
