Ecker András, Lázár Bence, Tóth Roland Imre, Urbán Martin, Tokodyné Szabadi Nikolett, Salinas Aponte Maria Teresa, Adnan Mohd, Várkonyi Eszter, Gócza Elen
Department of Animal Biotechnology, Institute of Genetics and Biotechnology, Hungarian University of Agriculture and Life Sciences, 2100 Gödöllő, Hungary.
Agribiotechnology and Precision Breeding for Food Security National Laboratory, 2100 Gödöllő, Hungary.
Life (Basel). 2023 Mar 23;13(4):867. doi: 10.3390/life13040867.
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.
最近，由于与体内技术相比成本更低且稳定性更高，体外基因保存技术得到了广泛应用。能够保存雌性特异性W染色体连锁基因的方法之一是原始生殖细胞（PGC）冷冻。PGC可通过从处于汉密尔顿-汉堡第14-16阶段的胚胎中采血分离得到。在我们的实验中，我们使用了两个新建立的特兰西瓦尼亚黑裸颈鸡细胞系和来自我们基因库的四个细胞系。在本研究中，我们比较了两种不同的冷冻培养基（FAM1和FAM2）。在冷冻前（BF）以及培养第0天、第1天和第7天解冻后，对PGC的细胞数量和活力进行了测量。我们使用RT-qPCR分析了PGC中生殖细胞特异性的鸡vasa同源物（CVH）表达谱。我们发现，在解冻后的第0天，用FAM2培养基冷冻的细胞系中的细胞数量显著高于用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更高，但差异不显著。冷冻也影响了用两种冷冻培养基处理的雄性细胞系中鸡vasa同源物基因的表达。
