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Abstract：
Recently, in vitro gene preservation has gained ground thanks to its lower cost and higher stability compared to in vivo techniques. One of the methods that can preserve female-specific W chromosome-linked genes is primordial germ cell (PGC) freezing. PGCs can be isolated from Hamburger-Hamilton stage 14-16 embryos via blood sampling. In our experiment, we used two newly established Black Transylvanian naked neck chicken cell lines and four cell lines from our gene bank. We compared two different freezing media (FAM1 and FAM2) in this study. The cell number and viability of the PGCs were measured before freezing (BF) and after thawing on Day 0, Day 1, and Day 7 of cultivation. We analyzed the germ cell-specific chicken vasa homologue (CVH) expression profile in PGCs using RT-qPCR. We found that on Day 0, immediately after thawing, the cell number in cell lines frozen with the FAM2 medium was significantly higher than in the FAM1-treated ones. On Day 1 and Day 7, the cell number and viability were also higher in most cell lines frozen with FAM2, but the difference was insignificant. The freezing also affected the chicken vasa homologue gene expression in male lines treated with both freezing media.
摘要：
最近,与体内技术相比，体外基因保存因其更低的成本和更高的稳定性而获得了应用。可以保存雌性特异性W染色体连锁基因的方法之一是原始生殖细胞（PGC）冷冻。PGCs可以通过血液取样从Hamburger-Hamilton阶段14-16胚胎中分离。在我们的实验中,我们使用了两个新建立的BlackTransvylvanian裸颈鸡细胞系和基因库中的四个细胞系。我们在这项研究中比较了两种不同的冷冻培养基(FAM1和FAM2)。在培养的第0天、第1天和第7天,在冷冻(BF)之前和解冻之后测量PGCs的细胞数量和活力。我们使用RT-qPCR分析了PGCs中生殖细胞特异性鸡vasa同源物（CVH）的表达谱。我们发现在第0天解冻后，用FAM2培养基冷冻的细胞系中的细胞数量显着高于FAM1处理的细胞系。在第1天和第7天,大多数用FAM2冷冻的细胞系中的细胞数量和活力也较高,但差异不显著。冷冻还影响了用两种冷冻培养基处理的雄性品系中的鸡vasa同源基因表达。
