| Analysis of Mitochondrial Transfer in Direct Co-cultures of Human Monocyte-derived Macrophages (MDM) and Mesenchymal Stem Cells (MSC)
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Author:
Date:
2017-05-05
[Abstract] Mesenchymal stem/stromal cells (MSC) are adult stem cells which have been shown to improve survival, enhance bacterial clearance and alleviate inflammation in pre-clinical models of acute respiratory distress syndrome (ARDS) and sepsis. These diseases are characterised by uncontrolled inflammation often underpinned by bacterial infection. The mechanisms of MSC immunomodulatory effects are not fully understood yet. We sought to investigate MSC cell contact-dependent communication with alveolar macrophages (AM), professional phagocytes which play an important role in the lung inflammatory responses and anti-bacterial defence. With the use of a basic direct co-culture system, confocal microscopy and flow cytometry we visualised and effectively quantified MSC mitochondrial transfer to AM ...
[摘要] 间充质干/基质细胞（MSC）是成年干细胞，已被证明可以改善急性呼吸窘迫综合征（ARDS）和败血症临床前模型中的存活，增强细菌清除和减轻炎症。这些疾病的特征在于通常受细菌感染支持的不受控制的炎症。 MSC免疫调节作用的机制尚未完全了解。我们试图研究与肺泡巨噬细胞（AM），在肺炎症反应和抗菌防御中起重要作用的专业吞噬细胞的MSC细胞接触依赖性通信。通过使用基本的直接共培养系统，共聚焦显微镜和流式细胞术，我们通过隧道纳米管（TNT）显现并有效量化了MSC线粒体转移到AM。为了模拟人类AM，在粒细胞巨噬细胞集落刺激因子（GM-CSF）存在下，从人供体血液中分离原代单核细胞并分化成巨噬细胞（单核细胞衍生的巨噬细胞，MDM），从而允许适应AM样表型（de Almeida等人，2000; Guilliams等人，2013）。人骨髓衍生的MSC用线粒体特异性荧光染色标记，广泛洗涤，以1:20（MSC / ...
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| Assessing Mitochondrial Transport via Cytoplasmic Nanotubular Bridges in Cells
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Author:
Date:
2015-08-05
[Abstract] This protocol aims to study intercellular transport of mitochondria, dynamic cellular organelles via tunnelling nanotubes (TNT), a cell membrane extension of cytoskeletal elements. The nanotubular bridges or the tunnelling nanotube highways are one of the emerging new cell-to-cell communication systems which mediates exchange of cellular materials, most importantly as in our observation, mitochondria. Mesenchymal stem cells (MSC) have been well studied to be endowed with a highly efficient intercellular mitochondrial donation ability and this property is now proven crucial to its functional role of rescue in cellular therapy.
[摘要] 此协议旨在研究线粒体，动态细胞器通过隧道纳米管（TNT），细胞骨架元素的细胞膜扩展的细胞间运输。 纳米管桥或隧道纳米管高速公路是新兴的新的细胞到细胞通信系统之一，其介导细胞材料的交换，最重要的是在我们的观察中，线粒体。 间充质干细胞（MSC）已经被充分研究以赋予高效的细胞间线粒体捐赠能力，并且这种性质现在被证明对于其在细胞治疗中的挽救的功能作用是至关重要的。
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| Fluorescence Measurement of Postharvest Physiological Deterioration (PPD) in Cassava Storage Roots
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Author:
Date:
2013-09-20
[Abstract] Cassava (Manihot esculenta Crantz) is a perennial root crop in the tropics. Within 24-72 hours of harvest the storage roots deteriorate rapidly, thereby necessitating their prompt processing or consumption. Postharvest physiological deterioration (PPD) of cassava storage roots is the result of a rapid oxidative burst, which leads to discoloration of the vascular tissues. The various fluorogenic probes available for in vivo reactive oxygen species (ROS) imaging could reveal complex spatial and temporal dynamics in plant tissues. Fluorescence measurement of PPD became widely used assay for ROS. Most of the ROS probes passively diffuse across cell membranes localize in the mitochondria, and exhibit fluorescence. Due to its high sensitivity to ROS and ease of loading and ...
[摘要] 木薯（ Manihot esculenta Crantz）是热带地区的多年生根作物。 在收获的24-72小时内，存储根快速劣化，从而迫使其迅速加工或消费。 木薯储存根的采后生理劣化（PPD）是快速氧化爆发的结果，其导致血管组织的变色。 可用于体内活性氧（ROS）成像的各种荧光探针可以揭示植物组织中的复杂空间和时间动力学。 PPD的荧光测量成为广泛使用的ROS测定法。 大多数ROS探针被动扩散穿过细胞膜定位在线粒体中，并且显示荧光。 由于其对ROS的高敏感性和易于加载和检测，二氢罗丹明123探针已广泛用于植物中以监测响应于各种刺激和发育过程范围的ROS积累。
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