从圈养的安的列斯海牛(Trichechus manatus manatus)皮肤活检中获得体细胞组织库和细胞培养的初步步骤。

IF 1.2 4区 生物学 Q3 VETERINARY SCIENCES Zoo Biology Pub Date : 2023-11-01 Epub Date: 2023-08-03 DOI:10.1002/zoo.21798
Matheus B Nascimento, Yasmin B F Moura, Radan E M Oliveira, Gabriela P O Lira, Alana A Borges, Fábia O Luna, Fernanda L N Attademo, Alexsandra F Pereira
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引用次数: 0

摘要

由于生态系统退化和日益严重的污染,安的列斯海牛的数量不断下降,这引起了人们对该物种保护策略的兴趣。在这种情况下,体细胞组织库是获取物种知识的重要工具,也是获得生物技术和生态毒理学应用的体细胞的重要工具。因此,本研究旨在探讨慢速冷冻(SF)和固体表面玻璃化(SSV)对圈养安的列斯海牛真皮组织超微结构和细胞活力的影响。虽然SSV没有改变真皮厚度,但SF保持了组织增殖潜力,通过核仁组织者区域面积来评估,类似于非冷冻保存的组织。此外,这两种技术都减少了成纤维细胞的数量,增加了胶原纤维的百分比。然而,只有用SF冷冻保存的组织和非冷冻保存的组织在体外培养后能够产生细胞。虽然SF没有改变细胞活力和增殖活性,但与非冷冻保存组织的细胞相比,来自冷冻保存组织的细胞表现出代谢降低、凋亡改变、活性氧水平升高和线粒体膜电位升高。综上所述,我们首次证明了SF可以低温保存安的列斯海牛体细胞组织,并在体外培养后获得细胞。为了提高该物种体细胞组织库的质量,需要改进体细胞样本的冷冻保存条件,特别是玻璃化。
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The initial steps toward the formation of somatic tissue banks and cell cultures derived from captive Antillean manatee (Trichechus manatus manatus) skin biopsies.

The declining population of the Antillean manatee caused by ecosystem degradation and rising pollution has prompted interest in developing conservation strategies for this species. Given this scenario, somatic tissue banks are important tools for acquiring knowledge about the species, as well as for obtaining somatic cells for biotechnological and ecotoxicological applications. Therefore, we aimed to assess the effects of slow freezing (SF) and solid-surface vitrification (SSV) of the dermis of captive Antillean manatees on the histology and ultrastructure of the tissue and cell viability in culture. While the SSV did not change the dermis thickness, the SF maintained the tissue proliferative potential, assessed by the nucleolar organizer region area, similar to noncryopreserved tissues. Moreover, both techniques reduced the number of fibroblasts and increased the percentage of collagen fibers. Nevertheless, only tissues cryopreserved with SF and noncryopreserved tissues were able to produce cells after in vitro culture. Although SF did not alter cell viability and proliferative activity, cells derived from cryopreserved tissues showed decreased metabolism, altered apoptosis, increased levels of reactive oxygen species, and mitochondrial membrane potential compared to cells from noncryopreserved tissues. In summary, we demonstrated for the first time that Antillean manatee somatic tissues can be cryopreserved by SF, and cells can be obtained after in vitro culture. Improvements in cryopreservation conditions, especially vitrification, of somatic samples are needed to increase the quality of somatic tissue banks in this species.

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来源期刊
Zoo Biology
Zoo Biology 生物-动物学
CiteScore
2.50
自引率
15.40%
发文量
85
审稿时长
6-12 weeks
期刊介绍: Zoo Biology is concerned with reproduction, demographics, genetics, behavior, medicine, husbandry, nutrition, conservation and all empirical aspects of the exhibition and maintenance of wild animals in wildlife parks, zoos, and aquariums. This diverse journal offers a forum for effectively communicating scientific findings, original ideas, and critical thinking related to the role of wildlife collections and their unique contribution to conservation.
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