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Crystal Violet

水晶紫

Company: Sigma-Aldrich
Catalog#: C3886
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VLA-4 Affinity Assay for Murine Bone Marrow-derived Hematopoietic Stem Cells
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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 - ...

Mouse Model of Dengue Virus Infection with Serotypes 1 and 2 Clinical Isolates
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; ...

Isolation and Culture of Human CD133+ Non-adherent Endothelial Forming Cells
Author:
Date:
2016-04-05
[Abstract]  Circulating endothelial progenitor cells (EPCs) have been the focus of many clinical trials due to their roles in revascularisation following ischemic events such as acute myocardial infarction as well as their contribution to vascular repair during organ transplantation. Research on EPCs has been controversial due to the lack of distinct markers expressed at the cell surface and varying methods for isolation and culture have resulted in the identification of a multitude of cell types, with differing phenotype and function, all falling under the label of “EPCs”. The most widely documented EPCs isolated for cell therapy are adherent in nature and lacking the progenitor markers such as CD133 and therefore unlikely to represent a true circulating EPC, the cells mobilised in response to a ... [摘要]  循环内皮祖细胞(EPCs)已经成为许多临床试验的焦点,因为它们在缺血事件例如急性心肌梗死后的血管再形成中的作用以及它们在器官移植期间对血管修复的贡献。由于缺乏在细胞表面表达的不同标记物,EPCs的研究已经引起争议,并且用于分离和培养的不同方法导致鉴定了具有不同表型和功能的多种细胞类型,所有这些都属于" EPCs"。分离用于细胞治疗的最广泛记载的EPCs本质上是粘附的,缺乏祖细胞标志物例如CD133,因此不太可能代表真正的循环EPC,所述细胞响应于血管损伤而动员。
我们最近发布了非粘附内皮形成细胞(naEFCs)群体的分离和广泛表征(Appleby等人,2012)(图1)。这些细胞与成熟内皮细胞标记(VEGFR2,CD144和CD31)一起表达祖细胞标记(CD133,CD34,CD117,CD90和CD38)。这些细胞还表达低水平的CD45,但不表达将其与"早期"EPC区分开的淋巴标志物(CD3,CD4,CD8)或骨髓标志物(CD11b和CD14),"晚期生长EPC"[最近称为内皮细胞集落形成细胞(ECFCs)]以及成熟内皮细胞(ECs)。图2A例示了naEFC的表面表达谱。功能研究证明这些naEFC(i)结合于玻璃体凝集素(图2A),(ii)显示乙酰化低密度脂蛋白摄取,(iii)增加血管细胞粘附分子(VCAM-1)响应于肿瘤坏死因子的表达和(iv)与成熟EC的共培养增加了在三维体外基质中的管,小管分支和环的数目。更重要的是,放置在体内的naEFC产生了包含由表达CD144的人EC内衬的脉管系统的新腔,并且有助于科学知识的各种进步(Appleby等人,2012; ...

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