Xu Hong-Yan, Hong Xiao-You, Zhong Chao-Yue, Wu Xu-Ling, Zhu Xin-Ping
Key Laboratory of Freshwater Fish Reproduction and Development (Ministry of Education), Key Laboratory of Aquatic Sciences of Chongqing, College of Fisheries, Southwest University, Chongqing 402460, China.
Key Laboratory of Tropical & Subtropical Fishery Resource Application & Cultivation of Ministry of Agriculture, Pearl River Fisheries Research Institute, Chinese Academy of Fishery Sciences, Guangzhou 510380, China.
Biology (Basel). 2022 May 22;11(5):790. doi: 10.3390/biology11050790.
Germ cells, as opposed to somatic cells, can transmit heredity information between generations. Cryopreservation and in vitro culture of germ cells are key techniques for genetic resource preservation and cellular engineering breeding. In this study, two types of cryopreserved samples, namely testis pieces and testicular cells of American shad, were comparatively analyzed for cell viability. The results showed that the cell viability of the cryopreserved testis pieces was much higher than that of the cryopreserved testicular cells. The viability of cells from the cryopreserved testis pieces ranged from 65.2 ± 2.2 (%) to 93.8 ± 0.6 (%), whereas the viability of the dissociated cells after cryopreservation was 38.5 ± 0.8 (%) to 87.1 ± 2.6 (%). Intriguingly, the testicular cells from the post-thaw testicular tissue could be cultured in vitro. Likewise, most of the cultured cells exhibited germ cell properties and highly expressed Vasa and PCNA protein. This study is the first attempt to effectively preserve and culture the male germ cells through freezing tissues in the American shad. The findings of this study would benefit further investigations on genetic resource preservation and other manipulations of germ cells in a commercially and ecologically important fish species.
与体细胞不同，生殖细胞能够在世代间传递遗传信息。生殖细胞的冷冻保存和体外培养是遗传资源保存和细胞工程育种的关键技术。在本研究中，对美洲西鲱的两种冷冻保存样本，即睾丸组织块和睾丸细胞，进行了细胞活力的比较分析。结果表明，冷冻保存的睾丸组织块的细胞活力远高于冷冻保存的睾丸细胞。冷冻保存的睾丸组织块的细胞活力范围为65.2±2.2（%）至93.8±0.6（%），而冷冻保存后解离细胞的活力为38.5±0.8（%）至87.1±2.6（%）。有趣的是，解冻后的睾丸组织中的睾丸细胞能够进行体外培养。同样，大多数培养细胞表现出生殖细胞特性，并高表达Vasa和PCNA蛋白。本研究是首次尝试通过冷冻组织有效保存和培养美洲西鲱的雄性生殖细胞。该研究结果将有助于对这种具有商业和生态重要性的鱼类的遗传资源保存及生殖细胞的其他操作进行进一步研究。
