Department of Biotechnology, Graduate School of Engineering, Nagoya University, Furo-cho, Chikusa-ku, Nagoya, 464-8603, Japan.
Cytotechnology. 2008 Jun;57(2):199-205. doi: 10.1007/s10616-008-9156-x. Epub 2008 Jul 23.
Primordial germ cells (PGCs) are precursors of germline cells. Although avian PGCs have been used to produce transgenic birds, their characteristics largely remain unknown. In this study, we isolated PGCs from chicken embryos at various developmental stages and analyzed the gene expression. Using the expression of stage-specific embryonic antigen-1 (SSEA-1) as a marker of chicken PGCs, we purified PGCs from embryos by fluorescence-activated cell sorting after incubation for 2.5-8.5 days. The number of SSEA-1(+) cells was almost unchanged during days 2.5-8.5 of incubation in females but continuously increased in male. Expression of several genes, including Blimp1, SOX2, and CXCR4, was observed in SSEA-1(+) cells but not in SSEA-1(-) cells in both female and male embryos. Quantitative reverse-transcription PCR analysis revealed that the expression of CXCR4, a chemokine receptor essential for migration of PGCs from the bloodstream to the gonads, was reduced after the circulating PGC stage (day 2.5).
原始生殖细胞（PGCs）是生殖细胞的前体细胞。尽管禽类 PGC 已被用于生产转基因鸟类，但它们的特性在很大程度上仍不清楚。在这项研究中，我们从不同发育阶段的鸡胚胎中分离出 PGC，并分析了它们的基因表达。使用阶段特异性胚胎抗原-1（SSEA-1）作为鸡 PGC 的标志物，我们在孵育 2.5-8.5 天后通过荧光激活细胞分选从胚胎中纯化 PGC。在雌性胚胎中，SSEA-1(+)细胞的数量在孵育的 2.5-8.5 天内几乎不变，但在雄性中则持续增加。在雌性和雄性胚胎中，SSEA-1(+)细胞中观察到几个基因的表达，包括 Blimp1、SOX2 和 CXCR4，但在 SSEA-1(-)细胞中没有。定量逆转录 PCR 分析显示，趋化因子受体 CXCR4 的表达在循环 PGC 阶段（第 2.5 天）后降低，该受体对于 PGC 从血液迁移到性腺至关重要。
