Efficient cryopreservation of Antillean manatee skin-derived somatic cells via reduced intracellular cryoprotectant concentration.

IF 1.5 4区 生物学 Q4 CELL BIOLOGY
Luanna Lorenna Vieira Rodrigues, Lhara Ricarliany Medeiros de Oliveira, Yasmin Beatriz França Moura, Yara Letícia Frutuoso E Silva, João Vitor da Silva Viana, Leonardo Vitorino Costa de Aquino, Radan Elvis Matias de Oliveira, Fernanda Loffler Niemeyer Attademo, Fábia Oliveira Luna, Alexsandra Fernandes Pereira
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Abstract

The declining population of the Antillean manatees has prompted interest in developing conservation strategies, including somatic cell cryopreservation. However, the type and concentration of intracellular cryoprotectant agents (CPAs) are limiting factors for its success. Therefore, we evaluated three concentrations (5, 8, 10%) of dimethyl sulfoxide (Me2SO) and ethylene glycol (EG) to assess if reducing CPA concentration is efficient for the cells of these animals. Cells not subjected to cryopreservation were used as a control. All cells were analyzed for morphology, viability, metabolism, proliferative activity (PDT), apoptosis, levels of reactive oxygen species (ROS), and mitochondrial membrane potential (ΔΨm). Regardless of the solution used, the cryopreservation did not change frozen-thawed cells' morphology, metabolism, and apoptosis levels compared to control group cells (p > 0.05). Immediately after thawing, cells derived from the 8% Me2SO group-maintained viability similar to the control; after in vitro culture of thawed cells, this positive response of viability was observed only in cells cryopreserved in solutions containing 5% and 8% CPA, regardless the type of CPA. Interestingly, cells frozen in 8% Me2SO showed a higher PDT value than the other groups (p < 0.05). Cells frozen with 10% EG showed higher ROS than the control group (p < 0.05). Additionally, regardless of the solution used, cryopreservation resulted in a change in ΔΨm. In summary, reducing the concentration of CPAs (5 and 8%) helps with somatic cell quality, regardless of the CPA type used in Antillean manatees.

通过降低细胞内冷冻保护剂浓度高效低温保存安的列斯海牛皮肤来源体细胞。
安的列斯海牛数量的下降引起了人们对发展保护策略的兴趣,包括体细胞冷冻保存。然而,细胞内冷冻保护剂(cpa)的类型和浓度是其成功的限制因素。因此,我们评估了三种浓度(5,8,10%)的二甲亚砜(Me2SO)和乙二醇(EG),以评估降低CPA浓度是否对这些动物的细胞有效。未冷冻保存的细胞作为对照。分析所有细胞的形态、活力、代谢、增殖活性(PDT)、凋亡、活性氧(ROS)水平和线粒体膜电位(ΔΨm)。无论使用何种溶液,与对照组细胞相比,冷冻保存未改变冷冻解冻细胞的形态、代谢和凋亡水平(p > 0.05)。解冻后,8% Me2SO组的细胞保持与对照组相似的活力;解冻后的细胞体外培养后,只有在含有5%和8% CPA的溶液中冷冻保存的细胞中,无论CPA的类型如何,都能观察到这种积极的活力反应。有趣的是,在8% Me2SO中冷冻的细胞显示出比其他组更高的PDT值(p
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来源期刊
CiteScore
3.70
自引率
4.80%
发文量
96
审稿时长
3 months
期刊介绍: In Vitro Cellular & Developmental Biology - Animal is a journal of the Society for In Vitro Biology (SIVB). Original manuscripts reporting results of research in cellular, molecular, and developmental biology that employ or are relevant to organs, tissue, tumors, and cells in vitro will be considered for publication. Topics covered include: Biotechnology; Cell and Tissue Models; Cell Growth/Differentiation/Apoptosis; Cellular Pathology/Virology; Cytokines/Growth Factors/Adhesion Factors; Establishment of Cell Lines; Signal Transduction; Stem Cells; Toxicology/Chemical Carcinogenesis; Product Applications.
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