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Surgical Gauze Sponge - 4"x 4" 12 Ply

Company: Dynarex
Catalog#: 3243
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Murine Bronchoalveolar Lavage
Author:
Date:
2017-05-20
[Abstract]  A basic Bronchoalveolar lavage (BAL) procedure in mouse is described here. Cells and fluids obtained from BAL can be analyzed by Hema3-staining, immunostaining, Fluorescence-activated cell sorting (FACS), PCR, bicinchoninic acid protein assay, enzyme-linked immunosorbent assay (ELISA), luminex assays, etc., to examine the immune cells, pathogens, proteins such as cytokines/chemokines, and the expression levels of inflammation-related and other genes in the cells. This will help to understand the underlying mechanisms of these lung diseases and develop specific and effective drugs. [摘要]  这里描述了小鼠中基本的支气管肺泡灌洗(BAL)程序。可以通过Hema3染色,免疫染色,荧光激活细胞分选(FACS),PCR,二金鸡宁酸蛋白测定,酶联免疫吸附测定(ELISA),luminex检测等来分析从BAL获得的细胞和液体。 / em>,以检查免疫细胞,病原体,蛋白质如细胞因子/趋化因子,以及细胞中炎症相关基因和其他基因的表达水平。这将有助于了解这些肺部疾病的潜在机制,并开发具体有效的药物。

背景 支气管肺泡灌洗(BAL)是通常用于诊断肺部疾病(包括肺癌)的简单且典型的方法(Daubeuf和Frossard,2012)。它用于采样肺组分,以确定肺中的蛋白质组成,免疫细胞和病原体。肺部慢性炎症在肺癌起始和进展中起关键作用。为了阐明肺肿瘤发生的炎症的潜在机制,我们的实验室使用了一种基本的BAL方案来确定肺部免疫反应(Qu等人,2015; Zhou等)。 ,2015; Sun等人,2016; Zhou等人,2017)。

Isolation of Murine Alveolar Type II Epithelial Cells
Author:
Date:
2017-05-20
[Abstract]  We have optimized a protocol for isolation of alveolar type II epithelial cells from mouse lung. Lung cell suspensions are prepared by intratracheal instillation of dispase and agarose followed by mechanical disaggregation of the lungs. Alveolar type II epithelial cells are purified from these lung cell suspensions through magnetic-based negative selection using a Biotin-antibody, Streptavidin-MicroBeads system. The purified alveolar type II epithelial cells can be cultured and maintained on fibronectin-coated plates in DMEM with 10% FBS. This protocol enables specific investigation of alveolar type II epithelial cells at molecular and cellular levels and provides an important tool to investigate in vitro the mechanisms underlying lung pathogenesis. [摘要]  我们优化了从小鼠肺分离肺泡II型上皮细胞的方案。通过气管内滴注分泌酶和琼脂糖,然后机械解聚肺来制备肺细胞悬浮液。通过使用生物素抗体Streptavidin-MicroBeads系统的磁性阴性选择,从这些肺细胞悬液中纯化肺泡II型上皮细胞。可以将纯化的肺泡II型上皮细胞培养并维持在含有10%FBS的DMEM中的纤连蛋白包被的平板上。该方案能够在分子和细胞水平上对肺泡II型上皮细胞进行特异性研究,并提供了一种重要的工具,用于在体外研究肺发病机制的机制。

背景 肺泡II型上皮细胞在肺泡完整性维持,表面活性蛋白合成和分泌中起关键作用,并防止细菌和病毒的肺部感染。最近使用小鼠肺癌模型的研究已经证明,肺泡II型上皮细胞是由化学致癌物质和致癌突变诱导的腺瘤/腺癌的关键细胞(Qu 等人,2015; Zhou > et al。,2015和2017)。为了进一步扩大我们对肺泡II型上皮细胞在体内肺发病机制中的作用的理解,需要分离肺泡II型上皮细胞以允许体外精确的机理分析, EM>。基于先前的研究(Corti等人,1996; Rice等人,2002),在我们的实验室中使用了一种修饰的方法来分离高度纯化的,可行的和可培养的来自小鼠的肺泡II型上皮细胞(Zhou等人,2015; Sun等人,2016)。

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