| Functional ex-vivo Imaging of Arterial Cellular Recruitment and Lipid Extravasation
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Author:
Date:
2017-06-20
[Abstract] The main purpose of this sophisticated and highly versatile method is to visualize and quantify structural vessel wall properties, cellular recruitment, and lipid/dextran extravasation under physiological conditions in living arteries. This will be of interest for a broad range of researchers within the field of inflammation, hypertension, atherosclerosis, and even the pharmaceutical industry. Currently, many researchers are using in vitro techniques to evaluate cellular recruitment, like transwell or flow chamber systems with cultured cells, with unclear physiological comparability. The here introduced method describes in detail the use of a sophisticated and flexible method to study arterial wall properties and leukocyte recruitment in fresh and viable murine carotid arteries ex ...
[摘要] 这种复杂和多功能方法的主要目的是在活体动物的生理条件下可视化和量化结构血管壁性质，细胞募集和脂质/葡聚糖外渗。这对于广泛的炎症，高血压，动脉粥样硬化甚至制药行业的研究人员都是有兴趣的。目前，许多研究人员正在使用体外技术来评估细胞募集，如具有培养细胞的transwell或流动室系统，具有不清楚的生理学可比性。这里介绍的方法详细描述了使用复杂而灵活的方法来研究在动脉血流条件下体外新鲜和活的鼠颈动脉中的动脉壁特性和白细胞募集。该模型模拟体内情况，并允许使用从两个不同供体分离的细胞和动脉（例如，野生型与特异性敲除）组合成一个实验，从而提供关于白细胞和/或内皮细胞特性的信息两个捐助者。因此，该模型可以被认为是复杂和侵入性体内研究的替代方案，例如对比实验。
【背景】该方法的核心是应用双光子激光扫描显微镜（TPLSM）来显示安装在动脉造影腔室中的体外颈动脉，其已显示模拟体内情况下存在的生理条件（Megens et al。，2007）。新鲜的动脉，在我们这种情况下是小鼠颈动脉，但该方法也适用于其他类似大小的血管（包括人类血管）（Bloksgaard等，2015），仔细提取并安装在使用细线的两个玻璃微量移液管的动脉造影室。该腔室应提供足够的空间以用显微镜物镜（优选为具有> ...
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| 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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| Aorta Ring Assay
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Author:
Date:
2016-07-05
[Abstract] Angiogenesis is the nature and pathological process of blood vessel growth from pre-existing vascular buds. It plays an important role in cancer and cardiovascular disease. The aorta ring assay is an approach to study angiogenesis. In this experiment, we used the aorta of rat as the study material, cleaned the surrounding tissue of aorta and cut it into 1 mm long rings. Next, the rings were cultured in growth factor-reduced matrigel polymerized at 37 °C. Angiogenesis was assessed at 7 days by using an inverted microscope platform.
[摘要] 血管生成是来自预先存在的血管芽的血管生长的性质和病理过程。 它在癌症和心血管疾病中起重要作用。 主动脉环测定是血管生成的一种方法。 在这个实验中，我们使用大鼠主动脉作为材料，清洁主动脉的周围组织，并将其切成1 mm长的环。 然后将环在37℃下聚合的生长因子减少的基质胶中培养。 通过使用倒置显微镜平台在7天评估血管发生。
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