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Safe 2020 Class II Biological Safety Cabinets

Company: Thermo Fisher Scientific
Catalog#: Safe 2020
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Human Endothelial Cell Spheroid-based Sprouting Angiogenesis Assay in Collagen
Author:
Date:
2018-09-05
[Abstract]  Angiogenesis, the formation of new blood vessels from pre-existing ones plays an important role during organ development, regeneration and tumor progression. The spheroid-based sprouting assay is a well-established and robust method to study the influence of genetic alterations or pharmacological compounds on capillary-like tube formation of primary cultured endothelial cells. A major advantage of this assay is the possibility to study angiogenesis in a 3D environment. Endothelial cells are cultured as hanging drops to form spheroids. Those spheroids are embedded into a collagen matrix and tube formation is analyzed 24 h later. By analyzing sprout number and sprout length the effects of genetic manipulation or drug treatment on angiogenesis can be investigated. [摘要]  血管生成,从先前存在的血管形成新血管在器官发育,再生和肿瘤进展中起重要作用。 基于球体的发芽测定法是一种成熟且稳健的方法,用于研究遗传改变或药理学化合物对原代培养的内皮细胞的毛细血管样管形成的影响。 该测定的主要优点是可以在3D环境中研究血管生成。 将内皮细胞培养为悬滴以形成球状体。 将这些球状体嵌入胶原基质中,24小时后分析管形成。 通过分析发芽数和发芽长度,可以研究遗传操作或药物治疗对血管生成的影响。

【背景】血管为器官提供氧气和营养。在不再满足局部需求的情况下,细胞分泌血管内皮生长因子(VEGF)以诱导新血管的形成。新的容器芽由一个由茎细胞牵引的前端细胞组成(Potente和Makinen,2017)。血管生成在生理条件下(例如,肌肉和脂肪组织的生长)以及病理条件(例如,伤口愈合,黄斑变性和肿瘤生长)发生。因此,非常需要破译协调血管生成的基本机制并测试干扰病理性血管生成的化合物。

基于球体的发芽试验由Thomas Korff博士和Hellmut Augustin博士在90年代后期开发(Korff和Augustin,1999),使研究人员能够快速研究药物或基因操作对发芽血管生成的影响。稳健的方式(Heiss et al。,2015)。基于球体的发芽测定的一个重要优点是分析3D环境中的芽形成。这促进内皮细胞之间的细胞 - ...

Infection of Soybean Plants with the Insect Bacterial Symbiont Burkholderia gladioli and Evaluation of Plant Fitness
Author:
Date:
2017-12-20
[Abstract]  To investigate the establishment and consequences of host-microbe interactions, it is important to develop controlled infection assays suitable for each system, as well as appropriate methods to evaluate successful infection and its associated effects. Here, we describe a procedure for bacterial inoculation of soybean plants, followed by the assessment of systemic infection and impact on plant fitness. Soybean (Glycine max) seedlings were mechanically wounded using a device that mimics insect herbivory and inoculated with known cell numbers of Burkholderia gladioli bacteria previously isolated from an insect host. The impact on the plants was evaluated by monitoring changes in height, time to flowering and chlorophyll content during plant development, and by quantifying ... [摘要]  为了研究宿主 - 微生物相互作用的建立和后果,开发适用于每个系统的受控感染测定法以及评估成功感染及其相关作用的适当方法是重要的。在这里,我们描述了大豆植物的细菌接种程序,然后评估全身感染和对植物健康的影响。使用模拟昆虫食草动物的装置对大豆(Glycine max)幼苗进行机械性伤害,并用先前从昆虫宿主分离的已知细胞数目的伯克霍尔德氏菌(B.coli)进行接种。通过监测植物发育过程中身高,开花时间和叶绿素含量的变化以及通过与用无菌水接种的植物相比量化种子产量来评估对植物的影响。使用定量PCR和荧光原位杂交(FISH)在来自发育植物的组织中检查细菌感染的存在和增殖。

微生物与不同的真核生物建立共生关系,对宿主的适应性有着深远的影响,从有益到不利(Frank,1997)。在许多情况下,这些协会是直接或间接的影响与其他生物,如潜在的替代主机相互作用。举例来说,植物,微生物和昆虫之间有许多三方相互作用,其中微生物共生体在不同宿主之间传播并影响相关生物体的生理或生态(Frago等人)。 ,2012; Gilbert et al。,2012)。在植物性的Lagriinae甲虫中,与来自植物致病性分支的细菌(Burkholderia gladioli)建立了共生伙伴关系,表明这种关联在三方相互作用的情况下发展。先前已经证明了从 Lagria villosa ...

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