Magalhães Lívia C, Bhat Maajid H, Freitas Jeferson L S, Melo Luciana M, Teixeira Dárcio I A, Pinto Luiz C A, Câmara Lília M C, Duarte José M B, Freitas Vicente J F
1 Laboratory of Physiology and Control of Reproduction, Faculty of Veterinary, Ceará State University , Fortaleza, Brazil .
2 Laboratory of Medical Immunology, Biomedicine Center, Ceará Federal University , Fortaleza, Brazil .
Biopreserv Biobank. 2017 Oct;15(5):463-468. doi: 10.1089/bio.2017.0060. Epub 2017 Sep 18.
The brown brocket deer Mazama gouazoubira is 1 of the 10 recognized brocket deer of the Neotropical region. Recently, this species has suffered a population decline due to current threats, mainly poaching and habitat loss. Several studies have shown that some endangered species can benefit from interspecies somatic cell nuclear transfer technology through the use of their somatic cells, such as the fibroblasts. Thus, the aim of this study was to verify the viability and the effect of cryopreservation on fibroblasts after several passages. For this purpose, fibroblast cells were cultured until passages 4, 7, and 10 (cultured control groups) and cryopreserved in cryotubes (frozen/warmed groups). The cellular viability, functionality, and percentage of cells undergoing necrosis and apoptosis were evaluated. The survival rates were always higher than 80% irrespective of the tested group, except for passage 10 in the frozen/warmed group. Population doubling time of cultured cells from passage 10 was significantly higher than that of passages 4 and 7, exhibiting low metabolic activity and a higher percentage of cells in initial apoptosis. In conclusion, the M. gouazoubira fibroblast-derived cell line provides an essential resource for further studies regarding reproductive biotechniques and is likely to be useful as an ex situ conservation strategy.
褐南美泽鹿（Mazama gouazoubira）是新热带地区10种公认的南美泽鹿之一。最近，由于当前的威胁，主要是偷猎和栖息地丧失，该物种的数量有所下降。几项研究表明，一些濒危物种可以通过使用其体细胞，如成纤维细胞，从种间体细胞核移植技术中受益。因此，本研究的目的是验证多次传代后成纤维细胞的活力以及冷冻保存的效果。为此，将成纤维细胞培养至第4、7和10代（培养对照组），并保存在冷冻管中（冷冻/解冻组）。评估细胞活力、功能以及发生坏死和凋亡的细胞百分比。无论测试组如何，存活率始终高于80%，冷冻/解冻组第10代除外。第10代培养细胞的群体倍增时间显著高于第4代和第7代，表现出低代谢活性和较高比例的初始凋亡细胞。总之，褐南美泽鹿成纤维细胞系为生殖生物技术的进一步研究提供了重要资源，并且很可能作为一种迁地保护策略发挥作用。
