Yousefi Taemeh Sara, Dehdilani Nima, Goshayeshi Lena, Kress Clémence, Rival-Gervier Sylvie, Montillet Guillaume, Ebrahimi Vishki Rouzbeh, Pain Bertrand, Dehghani Hesam
Stem Cell Biology and Regenerative Medicine Research Group, Research Institute of Biotechnology, Ferdowsi University of Mashhad, Mashhad, Iran.
Department of Medical Biochemistry and Microbiology, Biomedical Center (BMC), Uppsala University, Uppsala, Sweden.
Sci Rep. 2025 Apr 7;15(1):11858. doi: 10.1038/s41598-025-93777-w.
Efficient long-term cultivation of chicken primordial germ cells (cPGCs) is essential for various avian research and biotechnology applications. Our study aimed to address the challenge of inconsistent culture success by investigating strain-specific variations and optimizing culture conditions using two distinct media: Ovotransferrin-enriched medium (OTM) and chicken serum-supplemented medium (CSM). We demonstrated that each chicken strain has unique nutritional requirements, with Hubbard cPGCs thriving in OTM and Bovans cPGCs favoring CSM. This strain-specific variation was effective in derivation and proliferation rates and the expression of stem cell-specific markers such as POU5F3/OCT4 and NANOG. Furthermore, our study confirmed the sustained germ cell identity of long-term cultured cPGCs through the expression of DAZL, DDX4, and EMA1 germ cell markers. We also showed that cultured cPGCs retained their migratory abilities and transfectability, successfully generating G0 germline chimeras and G1 transgenic Bovans chickens. These findings highlight the importance of optimized culture conditions depending on the genotype to enhance the viability and genetic stability of cPGCs, paving the way for more effective genetic modifications and conservation strategies in avian species.
鸡原始生殖细胞（cPGCs）的高效长期培养对于各种禽类研究和生物技术应用至关重要。我们的研究旨在通过研究品系特异性差异并使用两种不同的培养基优化培养条件，即富含卵转铁蛋白的培养基（OTM）和添加鸡血清的培养基（CSM），来应对培养成功率不一致的挑战。我们证明，每个鸡品系都有独特的营养需求，哈伯德cPGCs在OTM中生长良好，博万斯cPGCs则更适合CSM。这种品系特异性差异在衍生率、增殖率以及干细胞特异性标志物如POU5F3/OCT4和NANOG的表达方面都很明显。此外，我们的研究通过DAZL、DDX4和EMA1生殖细胞标志物的表达，证实了长期培养的cPGCs的生殖细胞身份得以维持。我们还表明，培养的cPGCs保留了它们的迁移能力和转染能力，成功地产生了G0生殖系嵌合体和G1转基因博万斯鸡。这些发现突出了根据基因型优化培养条件对于提高cPGCs的活力和遗传稳定性的重要性，为禽类更有效的基因修饰和保护策略铺平了道路。
