The germplasm is a resource and tool for the conservation of genetic diversity in animals, including birds. Securing germplasm is limited in most bird species due to difficulties in semen collection and germ cell isolation, lack of germ cell-specific markers, and in vitro culture systems. Here, we report the production of germline chimeric quails by transplant of cryopreserved testicular cells (TCs) into the developing embryo. The testicular germ cell properties were maintained after freeze-thaw, with no significant reduction in cell viability irrespective of storage length. Cryopreserved TCs were transferred into Hamburger Hamilton (HH) stage 14-17 quail embryos, and were demonstrated to migrate into the embryonic gonads with similar efficiency to freshly isolated TCs. Twenty of 81 recipient embryos yielded hatchlings from cryopreserved TCs and the germline transmission efficiency was similar to that of freshly isolated cells. In conclusion, cryopreserved adult quail TCs are capable of (de)differentiation into functional gametes in recipient quail gonads and can generate donor TCs-derived progenies. This system is feasible for the isolation of sufficient germplasm resources from various bird species for conservation purposes.

中文翻译：

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通过将冷冻保存的睾丸细胞移植到发育中的胚胎中来生产种系嵌合鹌鹑

种质是保护动物（包括鸟类）遗传多样性的资源和工具。由于精液收集和生殖细胞分离困难、缺乏生殖细胞特异性标记物以及体外培养系统，大多数鸟类物种的种质保护受到限制。在这里，我们报告了通过将冷冻保存的睾丸细胞 (TC) 移植到发育中的胚胎中来生产种系嵌合鹌鹑。冻融后睾丸生殖细胞特性保持不变，无论储存时间长短，细胞活力都没有显着降低。冷冻保存的 TC 被转移到汉堡汉密尔顿 (HH) 14-17 期鹌鹑胚胎中，并被证明以与新鲜分离的 TC 相似的效率迁移到胚胎性腺中。81 个受体胚胎中有 20 个从冷冻保存的 TCs 中产生了幼体，生殖系传递效率与新鲜分离的细胞相似。总之，冷冻保存的成年鹌鹑 TCs 能够（去）分化为受体鹌鹑性腺中的功能配子，并且可以产生供体 TCs 衍生的后代。该系统对于从各种鸟类中分离出足够的种质资源用于保护目的是可行的。