Cardoso Camila Alampe, Motta Lina Castelo Branco, de Oliveira Vanessa Cristina, Martins Daniele Dos Santos
Faculdade de Zootecnia e Engenharia de Alimentos, Universidade de São Paulo, Pirassununga, SP, Brasil.
Hospital Veterinário, Universidade de Trás-os-Montes e Alto Douro, Vila Real, Portugal.
Anim Reprod. 2020 Sep 18;17(3):e20200044. doi: 10.1590/1984-3143-AR2020-0044.
The creation of a genetic resource bank of avian species aims to prevent the decline and fragmentation of wild bird populations, which in turn lead to the loss of genetic diversity and, in more serious cases, the extinction of the most threatened species. In order for the collected genetic material to be stored in a bank and useful when necessary, it is essential to improve the technique ensuring its effectiveness. Thus, our study used feather follicle cells from the species to standardize the technique of cell culture and subsequent cryopreservation. This study aimed to establish a protocol, , of isolation and primary culture of somatic cells derived from the feather follicle, with the purpose of establishing a cell lineage, and evaluate its viability for the biobank formation. Developing feathers of were collected at 12, 21 and 34 days of age. The feathers were morphologically analyzed and then we selected the region of the calamus due to the presence of pulp for cell culture and cryopreservation. The results showed that it is possible to find cells with distinct morphology; cells in elliptical shape with central nucleus also in elliptical shape, cells with shape and round nucleus, cells compatible with the fibers of the barbules, cell agglomerates and cells adhered to the bottom of the plate with fibroblastatoid shape. After 24 hours of culture there was the presence of primary culture with 80% of confluence and after cryopreservation the average viability after freezing was 68.8%, with cellular morphologies being maintained. Therefore, we proved the isolation of somatic cells from the follicle of bird's feathers, suggesting that this is a source of great value, viable and effective for obtaining biological material for the elaboration of a biobank.
建立鸟类遗传资源库旨在防止野生鸟类种群数量下降和碎片化,而这反过来又会导致遗传多样性丧失,在更严重的情况下,会导致最濒危物种灭绝。为了将收集到的遗传物质存储在库中并在必要时发挥作用,提高确保其有效性的技术至关重要。因此,我们的研究使用该物种的羽毛毛囊细胞来标准化细胞培养及后续冷冻保存技术。本研究旨在建立一套从羽毛毛囊中分离和原代培养体细胞的方案,目的是建立细胞系,并评估其用于生物样本库构建的可行性。在12日龄、21日龄和34日龄收集发育中的羽毛。对羽毛进行形态学分析,然后由于髓质的存在,我们选择羽根区域用于细胞培养和冷冻保存。结果表明,可以找到形态各异的细胞;椭圆形细胞,中央细胞核也是椭圆形,有特定形状和圆形细胞核的细胞,与羽小枝纤维相容的细胞,细胞聚集体以及成纤维细胞样形状且附着在培养皿底部的细胞。培养24小时后出现原代培养,汇合度达80%,冷冻保存后冷冻后的平均活力为68.8%,细胞形态得以维持。因此,我们证明了从鸟类羽毛毛囊中分离出体细胞,这表明这是一个具有巨大价值、可行且有效的获取生物材料以构建生物样本库的来源。