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从脂肪组织和脐带基质中分离的犬间充质干细胞的生物学特性和代谢特征。

Biological characteristics and metabolic profile of canine mesenchymal stem cells isolated from adipose tissue and umbilical cord matrix.

机构信息

Department of Veterinary Medical Sciences, University of Bologna, Ozzano Emilia, Bologna, Italy.

Health Science and Technologies Interdepartmental Center for Industrial Research (HST-ICIR), University of Bologna, Bologna, Italy.

出版信息

PLoS One. 2021 Mar 4;16(3):e0247567. doi: 10.1371/journal.pone.0247567. eCollection 2021.

Abstract

Despite the increasing demand of cellular therapies for dogs, little is known on the differences between adult and fetal adnexa canine mesenchymal stem cells (MSCs), and data on their metabolic features are lacking. The present study aimed at comparing the characteristics of canine adipose tissue (AT) and umbilical cord matrix (UC) MSCs. Moreover, for the first time in the dog, the cellular bioenergetics were investigated by evaluating the two main metabolic pathways (oxidative phosphorylation and glycolysis) of ATP production. Frozen-thawed samples were used for this study. No differences in mean cell proliferation were found (P>0.05). However, while AT-MSCs showed a progressive increase in doubling time over passages, UC-MSCs showed an initial post freezing-thawing latency. No differences in migration, spheroid formation ability, and differentiation potential were found (P>0.05). RT-PCR analysis confirmed the expression of CD90 and CD44, the lack of CD14 and weak expression of CD34, mostly by AT-MSCs. DLA-DRA1 and DLA-DQA1 were weakly expressed only at passage 0 by UC-MSCs, while they were expressed at different passages for AT-MSCs. There was no difference (P>0.05) in total ATP production between cell cultures, but the ratio between the "mitochondrial ATP Production Rate" and the "glycolytic ATP Production Rate" was higher (P<0.05) in AT- than in UC-MSCs. However, in both MSCs types the mitochondrial respiration was the main pathway of ATP production. Mitochondrial respiration and ATP turnover in UC-MSCs were higher (P<0.05) than in AT-MSCs, but both had a 100% coupling efficiency. These features and the possibility of increasing the oxygen consumption by a spare respiratory capacity of four (AT-MSCSs) and two (UC-MSCs) order of magnitude greater than basal respiration, can be taken as indicative of the cell propensity to differentiate. The findings may efficiently contribute to select the most appropriate MSCs, culture and experimental conditions for transplantation experiments in mesenchymal stem cell therapy for companion animals.

摘要

尽管对犬类细胞疗法的需求不断增加,但对于成年和胎儿附器犬间充质干细胞(MSCs)之间的差异知之甚少,并且缺乏有关其代谢特征的数据。本研究旨在比较犬脂肪组织(AT)和脐带基质(UC)MSCs 的特征。此外,这是首次在犬中通过评估产生 ATP 的两种主要代谢途径(氧化磷酸化和糖酵解)来研究细胞生物能学。本研究使用冷冻解冻样本。细胞增殖的平均值无差异(P>0.05)。然而,AT-MSCs 的倍增时间呈逐渐增加趋势,而 UC-MSCs 在冷冻解冻后有初始潜伏期。迁移、球体形成能力和分化潜力无差异(P>0.05)。RT-PCR 分析证实了 CD90 和 CD44 的表达,缺乏 CD14 和 CD34 的弱表达,主要由 AT-MSCs 表达。仅在 UC-MSCs 的传代 0 时,DLA-DRA1 和 DLA-DQA1 表达较弱,而在 AT-MSCs 中则在不同传代时表达。细胞培养物之间的总 ATP 产量无差异(P>0.05),但“线粒体 ATP 产生速率”与“糖酵解 ATP 产生速率”的比值在 AT-MSCs 中较高(P<0.05)。然而,在两种 MSC 类型中,线粒体呼吸都是 ATP 产生的主要途径。UC-MSCs 的线粒体呼吸和 ATP 周转率较高(P<0.05),但均具有 100%的偶联效率。这些特征以及通过比基础呼吸高四个(AT-MSCs)和两个(UC-MSCs)数量级的备用呼吸能力增加耗氧量的可能性,可以作为细胞分化倾向的指标。这些发现可以有效地帮助选择最适合的 MSC 、培养和实验条件,用于伴侣动物间充质干细胞治疗中的移植实验。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5d5f/7932077/8e28b73c8017/pone.0247567.g001.jpg

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