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Analysis of chicken primordial germ cells

机译:鸡原始生殖细胞分析

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摘要

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).
机译:原始生殖细胞(PGC)是生殖细胞的前体。尽管禽类PGC已被用于生产转基因禽类,但其特征在很大程度上仍然未知。在这项研究中,我们从不同发育阶段的鸡胚中分离了PGC,并分析了基因表达。使用阶段特异性胚胎抗原-1(SSEA-1)的表达作为鸡PGC的标记,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了PGC。在雌性孵化的2.5-8.5天中,SSEA-1 +细胞的数量几乎没有变化,而雄性则持续增加。在SSEA-1 +细胞中观察到了几种基因的表达,包括Blimp1,SOX2和CXCR4,而在SSEA-1中没有观察到。雌性和雄性胚胎中的细胞。定量逆转录PCR分析显示,循环PGC阶段(第2.5天)后,CXCR4(一种对PGCs从血流向性腺迁移所必需的趋化因子受体)的表达降低了。

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