Activation of Fibroblast Contractility via Cell-Cell Interactions and Soluble Signals
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Author:
Date:
2018-09-20
[Abstract] The collagen contraction assay is an in vitro, three-dimensional method to determine the factor(s) affecting the contractile behavior of activated cells such as fibroblasts in either physiological or pathological scenarios. The collagen lattices/hydrogels are seeded with fibroblasts to mimic the interactions between these cells and their surrounding extracellular matrix proteins in the connective tissue. This method is an important platform to assess components as potential therapeutic targets to prevent pathologies such as fibrosis, which are manifestations of hyperactivated fibroblasts. We have described a basic version of this collagen contraction assay, which is amenable to customization using different cell types under diverse experimental conditions.
[摘要] 胶原收缩测定是体外三维方法,用于确定影响生理或病理场景中活化细胞如成纤维细胞的收缩行为的因子。 胶原蛋白晶格/水凝胶用成纤维细胞接种,以模拟这些细胞与其周围细胞外基质蛋白在结缔组织中的相互作用。 该方法是评估组分作为潜在治疗靶标的重要平台,以预防纤维化等病症,这些病症是过度活化的成纤维细胞的表现。 我们已经描述了这种胶原收缩测定的基本版本,其适于在不同实验条件下使用不同细胞类型进行定制。
【背景】细胞外基质的组织收缩和重塑是许多生理条件(例如伤口愈合)中的基本过程。这两种现象的核心是成纤维细胞,它不仅产生和分泌细胞外基质蛋白,而且还可以通过机械相互作用重组它们。有趣的是,这些细胞行为通常在诸如纤维化的病理条件下被夸大(Desmoulière et al。,2005),从而说明需要理解这些过程的分子调节。虽然人们早就知道,胶原蛋白是细胞外基质的主要成分之一,是组织收缩的主要参与者(Bell et al。,1979),对机械细节的透彻理解。这个过程仍然难以捉摸。对体外成纤维细胞胶原基质体外收缩的研究使研究人员能够识别导致组织收缩的新型运动员(Ngo et al。,2006; Su and Chen, 2015年)。基于该测定,可溶性因子如TGFβ(Levi-Schaffer 等,1999)和免疫细胞(Garbuzenko et al。,2002; ...
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VLA-4 Affinity Assay for Murine Bone Marrow-derived Hematopoietic Stem Cells
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Author:
Date:
2017-02-20
[Abstract] Hematopoietic stem cells (HSCs) are defined by their functional ability to self-renew and to differentiate into all blood cell lineages. The majority of HSC reside in specific anatomical locations in the bone marrow (BM) microenvironment, in a quiescent non motile mode. Adhesion interactions between HSCs and their supporting BM microenvironment cells are critical for maintaining stem cell quiescence and protection from DNA damaging agents to prevent hematology failure and death. Multiple signaling proteins play a role in controlling retention and migration of bone marrow HSCs. Adhesion molecules are involved in both processes regulating hematopoiesis and stem- and progenitor-cell BM retention, migration and development. The mechanisms underlying the movement of stem cells from and to the ...
[摘要] 造血干细胞(HSC)由其自我更新的功能能力定义,并分化成所有血细胞谱系。大多数HSC位于骨髓(BM)微环境中的特定解剖位置,处于静止非运动模式。 HSCs与其支持的BM微环境细胞之间的粘附相互作用对于维持干细胞静止和保护免受DNA损伤因子阻止血液学失败和死亡至关重要。多重信号蛋白在控制骨髓HSCs的保留和迁移中起重要作用。粘附分子参与调节造血和干细胞和祖细胞BM保留,迁移和发育的两个过程。干细胞从骨髓移动到骨髓的机制尚未完全阐明,仍然是深入研究的对象。一个重要的方面是修饰干细胞保留,迁移和发育所需的干细胞和祖细胞的粘附分子的表达和亲和力。粘附力通过粘附分子的表达,亲和力和亲合力来调节。亲和力调节与分子结合识别和结合强度有关。在这里,我们描述了在我们的研究中使用的体外 FACS测定来研究整联蛋白α4亚类β1亚单位的表达,亲和力和功能也称为VLA-4)用于小鼠骨髓保留EPCR +长期复制HSC(LT-HSC)(Gur-Cohen等人,2015)。背景整合素是介导细胞和细胞 - ...
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Mouse Model of Dengue Virus Infection with Serotypes 1 and 2 Clinical Isolates
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Author:
Date:
2016-12-05
[Abstract] Dengue is a global public health threat caused by infection with any of the 4 related dengue virus serotypes (DENV1-4). Clinical manifestations range from self-limiting febrile illness, known as dengue fever (DF), to life-threatening severe diseases, such as dengue hemorrhagic fever (DHF) or dengue shock syndrome (DSS). Most cases of DHF/DSS are associated with secondary heterotypic infections through a phenomenon that is described as antibody-dependent enhancement of infection (ADE). There are an estimated 400 million human infections and several hundred thousand cases of severe dengue occurring yearly. At present, however, there are no approved antiviral drugs against DENV infection. The lack of a suitable animal model has hampered the evaluation of novel antiviral candidates for DENV ...
[摘要] 登革热是由4种相关登革热病毒血清型(DENV1-4)的任一种感染引起的全球公共卫生威胁。临床表现的范围从自限性发热疾病,称为登革热(DF),到危及生命的严重疾病,如登革热出血热(DHF)或登革热休克综合征(DSS)。大多数DHF/DSS病例通过描述为抗体依赖性增强感染(ADE)的现象与继发性异型感染相关。估计每年有4亿人感染和数十万例严重登革热病例。然而,目前,还没有批准的抗DENV感染的抗病毒药物。缺乏合适的动物模型阻碍了DENV感染的新抗病毒候选物的评价。由于DENV在免疫活性小鼠中不良地建立感染,已将AG129小鼠(缺乏I型和II型IFN [干扰素]受体)和小鼠适应的DENV2株应用于能够繁殖人类感染的几种主要病理的登革动物模型。最近,我们开发了具有临床分离株DENV1和DENV2的新的小鼠模型,其将用于药物测试和登革热发病机理研究(Watanabe等人,2016)。在这里我们描述建立临床分离株的登革热小鼠模型的细节;从体外材料制备到体内病毒感染。值得注意的是,由于DENV在小鼠中的感染性在病毒株之间不同,不是所有临床分离株都可以诱导严重的登革热。 关键字:登革热病毒,致命的小鼠模型,临床病毒,病毒感染的抗体依赖性增强,药物测试 [背景] 为了克服DENV在啮齿动物细胞中不能很好复制的缺点,多年来已经进行了许多努力来开发模拟人类登革热感染的小动物模型。近交小鼠模型系统允许实验可变性最小化,并且遗传工程小鼠模型能够再现动物中登革热临床症状的一些方面。过去的研究显示,用DENV2临床分离物感染的AG129小鼠(缺乏I型和II型IFN受体)感染瘫痪的迹象,这是中枢神经系统受累的病症,在人类病例中是罕见的(Shresta等人。,2004)。或者,产生可在AG129小鼠中诱导人类DHF/DSS样疾病的小鼠适应的DENV2毒株,并已用于登革热研究(Shresta等人,2006; ...
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