Institute of Biotechnology, National Taiwan University, Taipei 10672, Taiwan.
Univ Lyon, Université Lyon 1, Stem Cell and Brain Research Institute, U1208, USC1361, INSERM, INRA, Bron 69500, France.
Poult Sci. 2019 Apr 1;98(4):1820-1832. doi: 10.3382/ps/pey515.
This study aimed to isolate, culture, and characterize duck primordial germ cells (PGCs) and to compare these cells with chicken PGCs. We first cultured Muscovy duck (Cairina moschata) circulating PGCs and gonadal PGCs (gPGCs) in the modified serum-containing medium used to amplify chicken PGCs. gPGCs were found to proliferate better in serum-free chemically defined medium than in serum-containing medium. Thereafter, gPGCs were similarly isolated from 2 other duck breeds, the Pekin duck (Anas platyrhynchos) and the hybrid mule duck (C. moschata × A. platyrhynchos), and amplified for a limited period of time in the chemically defined culture condition, but sufficiently to be characterized and transplanted. Cultured gPGCs of all 3 duck breeds were characterized by Periodic acid-Schiff staining, immunocytochemical staining, and expression analysis of germline-specific and pluripotency genes. Cultured duck gPGCs colonized the gonads after being genetically labeled and injected into recipient embryos. Taken together, these results demonstrate that duck PGCs retain their germline characteristics after being isolated, expanded in vitro, and genetically modified. Further studies are required to establish the optimal conditions for long-term culture of duck PGCs, which may involve supplementing the culture medium with other growth factors or compounds.
本研究旨在分离、培养和鉴定鸭原始生殖细胞(PGCs),并将其与鸡 PGC 进行比较。我们首先在用于扩增鸡 PGC 的改良含血清培养基中培养麝香鸭(Cairina moschata)循环 PGC 和生殖腺 PGC(gPGCs)。发现 gPGCs 在无血清化学定义培养基中的增殖能力优于含血清培养基。此后,我们从另外 2 个鸭品种,即北京鸭(Anas platyrhynchos)和杂交骡鸭(C. moschata×A. platyrhynchos)中同样分离出 gPGCs,并在化学定义培养条件下进行有限时间的扩增,但足以进行鉴定和移植。用过碘酸希夫染色、免疫细胞化学染色和种系特异性和多能性基因的表达分析对所有 3 个鸭品种的培养 gPGCs 进行了鉴定。经基因标记并注射到受体胚胎中的培养鸭 gPGCs 定植于生殖腺。总之,这些结果表明,鸭 PGC 在分离、体外扩增和遗传修饰后保留其种系特征。需要进一步研究以确定鸭 PGC 长期培养的最佳条件,这可能涉及在培养基中添加其他生长因子或化合物。