A Simple Cryopreservation Method for Efficient Isolation of Live Cells from Dead Animals

IF 0.8 4区 生物学 Q3 ZOOLOGY Mammal Study Pub Date : 2022-02-02 DOI:10.3106/ms2021-0019
Michiya Sano, Ayako Kawanabe, Y. Kurosawa, Yusuke Suzuki, M. Takeda, Tomoaki Nakamura, H. Iwata, T. Kuwayama, K. Shirasuna
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引用次数: 1

Abstract

Abstract. Cryopreservation of somatic tissues and cells can be applied to biodiversity conservation. Although vitrification is widely used for tissue cryopreservation, it is challenging to obtain viable cells in facilities that lack adequate experimental tools, such as zoos. In this study, we established a simple tissue cryopreservation method for obtaining viable cells. Using mouse tissues of the ears and skin, we explored the conditions suitable for cryopreservation. After freezing, the tissues were thawed, and the cells were isolated. The tissues were then cut into small pieces to obtain viable cells. The use of a cryopreservative solution and freezing at –80°C increased the probability of obtaining viable cells. Viable cells were obtained and cultured even after the ear tissues were stored at room temperature for 24 h. Our method allowed primary cells to be isolated and cultured from ear tissues of dead animal. Further, we examined whether cells isolated from cryopreserved tissues could be studied in vitro. We found that treatment with lipopolysaccharides and Poly I:C increased the mRNA expression of pro-inflammatory cytokines in wild boar cells. These data suggest that the simple cryopreservation method developed here can be applied to biodiversity conservation and basic science studies of wild animal cells.
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一种从死亡动物体内高效分离活细胞的简单低温保存方法
摘要体细胞组织和细胞的冷冻保存可以应用于生物多样性保护。尽管玻璃化被广泛用于组织冷冻保存,但在动物园等缺乏足够实验工具的设施中获得活细胞是一项挑战。在本研究中,我们建立了一种简单的组织冷冻保存方法来获得活细胞。利用小鼠的耳朵和皮肤组织,我们探索了适合冷冻保存的条件。冷冻后,将组织解冻,并分离细胞。然后将组织切成小块以获得活细胞。冷冻保存溶液的使用和-80°C的冷冻增加了获得活细胞的可能性。即使在耳朵组织在室温下储存24小时后,也能获得活细胞并进行培养。我们的方法允许从死亡动物的耳朵组织中分离和培养原代细胞。此外,我们检查了从冷冻保存的组织中分离的细胞是否可以在体外进行研究。我们发现,用脂多糖和聚I:C处理增加了野猪细胞中促炎细胞因子的mRNA表达。这些数据表明,这里开发的简单冷冻保存方法可以应用于生物多样性保护和野生动物细胞的基础科学研究。
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来源期刊
Mammal Study
Mammal Study ZOOLOGY-
CiteScore
1.70
自引率
20.00%
发文量
23
审稿时长
>12 weeks
期刊介绍: Mammal Study is the official journal of the Mammal Society of Japan. It publishes original articles, short communications, and reviews on all aspects of mammalogy quarterly, written in English.
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