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InvitrogenTM DAPI

DAPI(4'',6-二脒基-2-苯基吲哚,二乳酸盐)

Company: Thermo Fisher Scientific
Catalog#: D3571
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Vascular Permeability Assay in Human Coronary and Mouse Brachiocephalic Arteries
Author:
Date:
2018-10-20
[Abstract]  Coronary artery disease remains an important cause of morbidity and mortality. Previous work, including ours, has focused on the role of intraplaque hemorrhage, particularly from immature microvessel angiogenesis, as an important contributor to plaque progression via increases in vascular permeability leading to further intraplaque hemorrhage, which increases red cell membrane-derived free cholesterol in plaque content and inflammatory cell recruitment. Evans Blue Dye (EBD) assay is widely used as a standard assay for vasculature permeability. However, the method has not been established in fresh human coronary artery autopsy samples to evaluate intraplaque microvessel permeability and angiogenesis. In this protocol, we describe a method to evaluate human coronary samples for ... [摘要]  冠状动脉疾病仍然是发病率和死亡率的重要原因。以前的研究,包括我们的研究,都集中在斑块内出血的作用,特别是来自未成熟的微血管血管生成,作为斑块进展的重要因素,通过增加血管通透性导致进一步的斑块内出血,增加斑块中红细胞膜来源的游离胆固醇内容和炎症细胞募集。 Evans Blue Dye(EBD)测定法广泛用作脉管系统渗透性的标准测定法。然而,该方法尚未在新鲜人冠状动脉尸检样本中建立,以评估斑块内微血管通透性和血管生成。在该方案中,我们描述了评估人类冠状动脉样本的微血管通透性的方法,包括灌注冠状动脉的程序,用于组织学分析和免疫染色的动脉样本的收集以及使用适当的方法来分析图像。还提供了在小鼠模型中使用FITC-葡聚糖以评估血管通透性的任选程序。这些Evans Blue Dye程序可用于在各种病理条件下提供人样品和动物模型中内皮完整性和渗透性的功能测量。

【背景】
血管内皮细胞主动调节血浆成分和循环细胞(包括白细胞)从血液到亚内皮组织的浸润。这种机制通常被认为是动脉粥样硬化起始和发展的关键步骤(Mundi et al。>,2018)。血管通透性的调节通过内皮细胞 - 细胞连接的协调打开和闭合来实现。在几种疾病状态下,内源性药物如组胺,凝血酶和血管内皮生长因子(VEGF)显着但可逆地以不同方式改变细胞 - ...

Murine Hair Follicle Derived Stem Cell Transplantation onto the Cornea Using a Fibrin Carrier
Author:
Date:
2018-05-20
[Abstract]  The goal of this protocol is to establish a procedure for cultivating stem cells on a fibrin carrier to allow for eventual transplantation to the eye. The ability to transfer stem cells to a patient is critical for treatment for a variety of disorders and wound repair. We took hair follicle stem cells from the vibrissae of transgenic mice expressing a dual reporter gene under the control of the Tet-on system and the keratin 12 promoter (Meyer-Blazejewska et al., 2011). A clonal growth assay was performed to enrich for stem cells. Once holoclones formed they were transferred onto a fibrin carrier and cultivated to obtain a confluent epithelial cell layer. Limbal stem cell deficient (LSCD) mice were used as the transplant recipient in order to test for successful grafting and ... [摘要]  该方案的目标是建立一种在纤维蛋白载体上培养干细胞以允许最终移植到眼睛的程序。 将干细胞转移给患者的能力对于治疗各种疾病和伤口修复至关重要。 我们在Tet-on系统和角蛋白12启动子(Meyer-Blazejewska等人,2011)的控制下从表达双报告基因的转基因小鼠的触须中取出毛囊干细胞。 进行克隆生长测定以富集干细胞。 一旦形成holoclones,将它们转移到纤维蛋白载体上并培养以获得融合上皮细胞层。 将角膜缘干细胞缺陷(LSCD)小鼠用作移植受体以测试移植成功和最终分化为角膜上皮表型。

【背景】干细胞被广泛用作治疗工具,因此递送手段是必不可少的。实际上,许多研究人员和公司正在寻找将细胞输送到人体内以优化细胞存活以及整合到宿主组织中的最佳方式。注射方法已广泛用于动物模型,但往往导致生存和整合差。目前正在使用利用生物材料和手术装置的技术。一种用于输送干细胞的技术是纤维蛋白载体。纤维蛋白凝胶是可降解的生物聚合物,其可粘附于允许细胞附着,迁移和增殖的天然组织(Ehrbar et al。,2005)。纤维蛋白凝胶具有许多优点,包括生物相容性,受控降解(Kjaergard等人,1994; Sidelmann等人,2000),均匀细胞分布和高细胞接种效率(Swartz ,2005)。已经将纤维蛋白凝胶用于治疗皮肤烧伤(Pellegrini等人,1999; ...

Adhesion of Enteroaggregative E. coli Strains to HEK293 Cells
Author:
Date:
2018-04-20
[Abstract]  Enteroaggregative Escherichia coli (EAEC) is a recognized cause of acute diarrhea among both children and adults worldwide. EAEC strains are characterized by the presence of aggregative adherence fimbriae (AAF), which play a key role in pathogenesis by mediating attachment to the intestinal mucosa and by triggering host inflammatory responses. The aggregative adherence fimbria II (AAF/II) is the most important adherence factor of EAEC prototype strain 042 (EAEC042) to intestinal cells. Multiple receptors for AAF/II on epithelial cells have been identified including the transmembrane signaling mucin Muc1. This protocol describes a method to measure adherence of EAEC strains to HEK293 cells expressing the Muc1 glycoprotein. [摘要]  肠道集聚性大肠杆菌(EAEC)是全球儿童和成人急性腹泻的公认原因。 EAEC菌株的特征在于存在聚集粘附菌毛(AAF),其通过介导与肠粘膜的附着和通过引发宿主炎症反应而在发病机制中起关键作用。 聚合粘附菌毛II(AAF / II)是EAEC原型菌株042(EAEC042)对肠细胞最重要的粘附因子。 已经鉴定了上皮细胞上AAF / II的多种受体,包括跨膜信号传导粘蛋白Muc1。 该协议描述了测量EAEC菌株对表达Muc1糖蛋白的HEK293细胞的依从性的方法。

【背景】EAEC是世界范围内地方性和流行性腹泻病的重要原因。尽管发展中国家儿童腹泻最常见,但EAEC还与免疫受损成人腹泻,旅行者和工业化国家的食源性疾病有关,例如由志贺毒素(Stx)2a型产生的大致致命爆发2011年在北欧的血清型O104:H4的EAEC菌株(Harrington等人,2006; Rasko等人,2011)。 EAEC发病机制由生物体粘附肠细胞,产生肠毒素和细胞毒素并最终诱导炎症的能力决定(Harrington等,2006)。 EAEC对肠细胞的依从性由AAF菌毛粘附素介导(Czeczulin等人,1997)。迄今为止,已经描述了至少5种AAF菌毛的变体,全部编码在范围为55至65MDa的毒力质粒中(Jonsson等人,2015)。 ...

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