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Mitochondrial energetics and contents evaluated by flow cytometry in human maternal and umbilical cord blood.
Scandinavian Journal of Clinical and Laboratory Investigation ( IF 1.3 ) Pub Date : 2020-05-29 , DOI: 10.1080/00365513.2020.1768584
Julie Kristine Guldberg Stryhn 1, 2, 3 , Jacob Larsen 2, 4 , Palle Lyngsie Pedersen 2, 5 , Anne-Dorthe Feldthusen 2 , Jan Kvetny 2 , Peter Haulund Gæde 3, 6
Affiliation  

Background: Mitochondrial dysfunction may relate to metabolic disorders. The relation between maternal and fetal mitochondrial function needs attention due to heritage.

Objectives: To evaluate the use of the staining methods TetraMethylRhodamine Methyl Ester (TMRM) and Mitotracker Green (MTG) for flow cytometric measurements of umbilical cord blood mitochondrial function.

Methods: 53 euthyroid at-term pregnant women and their offspring were included by blood collections. The offspring had blood drawn from the clamped umbilical cord. Flow cytometry with MTG, TMRM and Propidium Iodide were performed the following day. A cell count (antibody coating and flow cytometry) was performed for 9 maternal and cord samples. As a quality control, blood of 32 healthy donors was evaluated by flow cytometric analyzes same day as sampling and the following day to test stability of the measurements.

Results: Cord mitochondrial measurements were lower than maternal. Maternal and cord mitochondrial function were positively correlated, especially reflected by MTG fluorescence-intensity (FI). Samples stored presented with very changed fluorescence patterns. However, the fluorescence intensity ratios MTG/TMRM of stained white blood cells were related within same day measurements, depicting an extensive and common bioenergetic cellular change.

Conclusion: Cord blood flow cytometry by MTG- and TMRM- staining is possible with fluorescence intensity positively correlated to maternal fluorescence intensity. Storage of blood triggers mitochondrial dynamics. The methods are applicable with certain reservations, and they benefit from their non-invasive character compared to mitochondrial evaluation by muscle-biopsies.



中文翻译:

通过流式细胞术评估人母体和脐带血中的线粒体能量和含量。

背景:线粒体功能障碍可能与代谢异常有关。母体和胎儿线粒体功能之间的关系由于传统而需要引起重视。

目的:评估四甲基罗丹明甲基酯(TMRM)和线粒体绿(MTG)染色法在脐血线粒体功能流式细胞仪检测中的应用。

方法:对53例甲状腺功能正常的孕妇及其后代进行采血。后代的血液是从脐带钳中抽出的。第二天用MTG,TMRM和碘化丙啶进行流式细胞术。对9个母体和脐带样品进行细胞计数(抗体包被和流式细胞术)。作为质量控制,在采样当天和第二天通过流式细胞仪分析评估了32位健康供体的血液,以测试测量的稳定性。

结果:脐带线粒体测量值低于母亲。母体和脐带线粒体功能呈正相关,尤其是MTG荧光强度(FI)反映出来。储存的样品呈现出非常变化的荧光模式。然而,染白血细胞的荧光强度比MTG / TMRM在同一天的测量中是相关的,描述了广泛而常见的生物能细胞变化。

结论:通过MTG-和TMRM-染色进行脐血流式细胞术是可行的,其荧光强度与母体荧光强度呈正相关。血液储存会触发线粒体动力学。该方法在一定程度上适用,并且与通过肌肉活检进行的线粒体评估相比,得益于其非侵入性特征。

更新日期:2020-05-29
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