Name: harvesting murine alveolar macrophages and evaluating cellular activation induced by polyanhydride nanoparticles
Uploaded: 2012-06-08T12:00:00
Description: Watch this Scientific Journal Video about Harvesting Murine Alveolar Macrophages and Evaluating Cellular Activation Induced by Polyanhydride Nanoparticles at JoVE.com

作者想感谢来自衣阿华州立大学流式细胞设施美国陆军医学研究和装备司令部（格兰特W81XWH-09-1-0386和W81XWH-10-1-0806号）的财政支持和肖恩·里格比博士他的专家技术援助。

1。鼠标采用肺灌洗收获（AMɸ）肺泡巨噬细胞<ol><li>穿白大褂，一次性手套，和适当的保护眼睛，如适当的个人防护装备（PPE）。 </li><li>开始收获前准备完整AMɸ（cAMɸ）培养基。新增2.5青霉素/链霉素（笔/链球菌）解决方案，250β-巯基乙醇微升（巯基乙醇）和25热灭活胎牛血清（FBS），毫升222.25 Dulbelcco的修改鹰培养基（DMEM培养基）毫升毫升。过滤消毒cAMɸ介质在真空条件下使用0.2微米的孔大小瓶...

<ol>
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酸酐纳米疫苗平台显示效果时，给予单剂量疗法5滴鼻。在此疫苗交付平台引起的肺部测量驻地吞噬细胞群的激活，允许评估其潜在的能力，最终促进适应性免疫反应。 具体来说，从肺灌洗液中收获肺泡巨噬细胞和不同配方的纳米粒子治疗提供了不同的粒子化学能力的见解，激活巨噬细胞，导致抗原提呈6,8。此外， 在体外研究中，这些都是用于?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td> CAM媒体 </td><td></td><td></td><td></td></tr><tr><td> DMEM培养液</td><td> cellgro </td><td> 15-013-CV </td><td></td></tr><tr><td> 50毫米2 - 巯基乙醇</td><td>西格玛</td><td> M3148-25ML </td><td></td></tr><tr><td>青霉素/ 10000微克/毫升溶液</td><td> cellgro </td><td> 30-002-CI </td><td></td></tr><tr><td>胎牛血清</td><td>亚特兰大生物</td><td> S11150 </td><td></td></tr><tr><td> 流式细胞仪缓冲 </td><td></td><td></td><td></td></tr><tr><td>氯化钠</td><td> Fisher Scientific则</td><td> S671-500 </td><td></td></tr><tr><td>磷酸钠</td><td> Fisher Scientific则</td><td> MK7868500 </td><td></td></tr><tr><td>氯化钾</td><td> Fisher Scientific则</td><td> P217500 </td><td></td></tr><tr><td>磷酸钾</td><td> Fisher Scientific则</td><td> P288-200 </td><td></td></tr><tr><td>牛血清白蛋白（牛血清白蛋白） </td><td>西格玛</td><td> A7888 </td><td></td></tr><tr><td>叠氮化钠</td><td>西格玛</td><td> S2002 </td><td></td></tr><tr><td> 抗体 </td><td></td><td></td><td></td></tr><tr><td>鼠IgG </td><td>西格玛</td><td> I4341 </td><td></td></tr><tr><td>反女士CD16/32 </td><td> eBioscience </td><td> 16-0161 </td><td></td></tr><tr><td>抗MHC II类女士单体IA / IE浏览器，M5/114.15.2克隆，共轭异硫氰酸荧光素（FITC标记） </td><td> EBIOS新增5Y科学</td><td> 11-5321 </td><td></td></tr><tr><td>抗CD86的鼠标，克隆GL-1，共轭变藻蓝蛋白（APC）Cy7 </td><td> Biolegend </td><td> 105030 </td><td></td></tr><tr><td>抗小鼠CD40，克隆1C10，结合到APC </td><td> eBioscience </td><td> 17-0401 </td><td></td></tr><tr><td>抗鼠CD209，克隆5H10，结合生物素</td><td> eBioscience </td><td> 13-2091 </td><td></td></tr><tr><td>抗小鼠CD11b的克隆M1/70，共轭的Alexa Fluor 700 </td><td> eBioscience </td><td> 56-0112 </td><td></td></tr><tr><td> F4/80的反鼠标，克隆BM8，结合藻红蛋白（PE）Cy7 </td><td> eBioscience </td><td> 25-4801 </td><td></td></tr><tr><td> PE得克萨斯红色标记的链霉</td><td> BD公司</td><td> 551487 </td><td></td></tr><tr><td> 其他耗材和试剂 </td><td></td><td></td><td></td></tr><tr><td>乙醇</td><td> Fisher Scientific则</td><td> A405-20 </td><td>作为70％（V / V） </td></tr><tr><td>压缩的CO 2 </td><td> linweld </td><td> 16000060 </td><td></td></tr><tr><td> 1毫升注射器</td><td> BD公司</td><td> 309659 </td><td></td></tr><tr><td>主权3½“神父汤姆Catcatheter的</td><td>肯德尔</td><td> 703021 </td><td></td></tr><tr><td>生物安全柜</td><td> NUAIRE </td><td>系列22 </td><td></td></tr><tr><td>解剖剪刀</td><td>费舍尔</td><td> 138082 </td><td></td></tr><tr><td>镊子</td><td> roboz </td><td> RS-8254 </td><td></td></tr><tr><td> PBS，1X无钙，镁</td><td> cellgro </td><td> 21-040-CM </td><td></td></tr><tr><td> 15 mL离心管，带螺旋盖</td><td> VWR国际</td><td> 21008-216 </td><td></td></tr><tr><td>治疗板六以及组织文化</td><td> COSTAR </td><td> 3516 </td><td></td></tr><tr><td>塑料管架</td><td> NALGENE </td><td> 5970 </td><td></td></tr><tr><td>细胞刮刀24厘米</td><td>卟啉</td><td> 99002 </td><td></td></tr><tr><td> 5 mL聚苯乙烯回合中筒底部</td><td>鹘</td><td> 352008 </td><td></td></tr><tr><td>吸管，加大援助</td><td>德拉蒙德</td><td> 4-000-105 </td><td></td></tr><tr><td> 10，5，2毫升移液器</td><td>费舍尔</td><td> 13-675 </td><td></td></tr><tr><td> 200和10μL的micropipettors </td><td>吉尔森Pipetman </td><td> F123601 </td><td></td></tr><tr><td> 200和10μL枪头</td><td>费舍尔</td><td> 02-707 </td><td></td></tr><tr><td>屋宇署稳定固色剂</td><td> BD公司</td><td> 338036 </td><td></td></tr><tr><td> isoton第二稀释剂</td><td>贝克曼 - 库尔特</td><td> 8546719 </td><td></td></tr><tr><td> ZAP-oglobin II裂解试剂</td><td>贝克曼 - 库尔特</td><td> 7546138 </td><td></td></tr><tr><td>库尔特计数器的聚苯乙烯瓶</td><td>贝克曼 - 库尔特</td><td> 14310-684 </td><td></td></tr><tr><td>试管</td><td> BD公司</td><td> 352008 </td><td></td></tr><tr><td> 设备 </td><td></td><td></td><td></td></tr><tr><td>冷冻离心机</td><td> labnet </td><td> 50075040 </td><td></td></tr><tr><td>湿润的孵化器的CO 2 </td><td> nuaire </td><td>模型自动流8500 </td><td></td></tr><tr><td> FACSCanto流式细胞仪</td><td> BD公司</td><td> 338960 </td><td></td></tr><tr><td>库尔特颗粒计数器Z1的</td><td>贝克曼 - 库尔特</td><td> WS-Z1DUALPC的</td><td></td></tr><tr><td> sonicator液体微尖加工设备</td><td> misonix </td><td>型号的S-4000 </td><td></td></tr></tbody></table>

harvesting murine alveolar macrophages and evaluating cellular activation induced by polyanhydride nanoparticles

可生物降解的纳米粒子已经出现作为一个通用平台的设计和实施新鼻内疫苗对呼吸道传染病。具体来说，酸酐纳米脂肪癸二酸（SA），芳香1,6 -双（P-羧基苯氧）正己烷（CPH），或双亲1,8 -双（P-羧基苯氧）-3,6 - dioxaoctane组成（CPTEG）显示独特的体积和表面侵蚀动力学1,2和可利用缓慢释放功能的生物大分子（如蛋白抗原，免疫球蛋白等） 在体内 3,4,5。这些纳米粒子也具有内在的辅助活动，使他们6,7,8疫苗交付平台的一个很好的选择。 为了阐明机制激活先天免疫后鼻腔粘膜接种，必须评估的分子和细胞反应的抗原P怨恨细胞（APC）负责启动免疫反应。树突状细胞是进行气管中发现的主要装甲运兵车，而在肺实质9,10,11（AMɸ）肺泡巨噬细胞为主。 AMɸ是非常有效地清除病原微生物和细胞碎片12,13肺部。此外，这种细胞类型发挥宝贵的作用在运输微生物抗原的淋巴结，这是一个重要的第一步启动适应性免疫反应9。 AMɸ也表达水平升高的先天模式识别和清道夫受体，分泌促炎介质，和总理幼稚T细胞12,14。一个人口相对纯的AMɸ（例如，大于80％），可以很容易地获得通过肺灌洗在实验室中研究。从免疫动物AMɸ收获的居民提供了一个代表性的表型的巨噬细胞，将encounter的粒子在体内的疫苗。在此，我们描述了从小鼠到收获和文化AMɸ使用的协议和检查酸酐纳米粒子在体外治疗后的巨噬细胞的激活型。

Watch this Scientific Journal Video about Harvesting Murine Alveolar Macrophages and Evaluating Cellular Activation Induced by Polyanhydride Nanoparticles at JoVE.com

Harvesting Murine Alveolar Macrophages and Evaluating Cellular Activation Induced by Polyanhydride Nanoparticles

在此，我们描述了小鼠肺泡巨噬细胞，居住在肺先天免疫细胞，并检查其激活，共培养酸酐纳米收获的协议。

收获小鼠肺泡巨噬细胞和评价聚酸酐纳米粒子诱导的细胞活化

Biodegradable nanoparticles have emerged as a  versatile platform for the design and implementation of new intranasal vaccines  against respiratory ...

Research

JoVE Journal

Bioengineering

Analyzing Cellular Internalization of Nanoparticles and Bacteria by Multi-spectral Imaging Flow Cytometry

由多光谱成像流式细胞仪分析细胞内化纳米粒子和细菌

Nanoparticulate  systems have emerged as valuable tools in vaccine delivery through their  ability to efficiently deliver cargo, including proteins, to ...

科研

生物工程

High-throughput Synthesis of Carbohydrates and Functionalization of Polyanhydride Nanoparticles

碳水化合物和聚酸酐纳米粒子功能化的高通量合成

Transdisciplinary approaches involving areas such as material design, nanotechnology, chemistry, and immunology have to be utilized to rationally design ...

Evaluation of Polymeric Gene Delivery Nanoparticles by Nanoparticle Tracking Analysis and High-throughput Flow Cytometry

高分子基因传递纳米粒子的纳米粒子跟踪分析和高通量流式细胞仪评价

Non-viral gene delivery using polymeric nanoparticles has emerged as an attractive approach for gene therapy to treat genetic diseases1 and as a ...

PLGA Nanoparticles Formed by Single- or Double-emulsion with Vitamin E-TPGS

Poly(lactic-co-glycolic acid) (PLGA) is a biocompatible member of the aliphatic polyester family of biodegradable polymers. PLGA has long been a popular ...

Using Cell-substrate Impedance and Live Cell Imaging to Measure Real-time Changes in Cellular Adhesion and De-adhesion Induced by Matrix Modification

使用Cell-基板阻抗和活细胞成像，以衡量矩阵修改诱导实时变化的细胞粘附和去粘附

Cell-matrix adhesion plays a key role in controlling cell morphology and signaling. Stimuli that disrupt cell-matrix adhesion (e.g., myeloperoxidase and ...

Microbiologically Induced Calcite Precipitation Mediated by Sporosarcina pasteurii

Microbiologically Induced Calcite Precipitation Mediated by Sporosarcina pasteurii

微生物诱导方解石沉淀通过介导<em&gt; Sporosarcina巴斯德</em

The particular bacterium under investigation here (S. pasteurii) is unique in its ability, under the right conditions, to induce the hydrolysis of urea ...

Identification of Metal Oxide Nanoparticles in Histological Samples by Enhanced Darkfield Microscopy and Hyperspectral Mapping

金属氧化物纳米颗粒的组织学样品鉴定增强暗场显微镜和高光​​谱遥感制图

Nanomaterials are increasingly prevalent throughout industry, manufacturing, and biomedical research. The need for tools and techniques that aid in the ...

Production of Metal Nanoparticles by Pulsed Laser-ablation in Liquids: A Tool for Studying the Antibacterial Properties of Nanoparticles

液体中脉冲激光烧蚀制备金属纳米粒子：研究纳米粒子抗菌性能的工具

The emergence of multidrug-resistant bacteria is a global clinical concern leading some to speculate about our return to a "pre-antibiotics" era of ...

Surface Functionalization of Hepatitis E Virus Nanoparticles Using Chemical Conjugation Methods

化学共轭法在戊型肝炎病毒纳米粒子表面功能化中的应用

Virus-like particles (VLPs) have been used as nanocarriers to display foreign epitopes and/or deliver small molecules in the detection and treatment of ...

Repressing Gene Transcription by Redirecting Cellular Machinery with Chemical Epigenetic Modifiers

用化学表观修饰因子重定向细胞机械抑制基因转录

Regulation of chromatin compaction is an important process that governs gene expression in higher eukaryotes. Although chromatin compaction and gene ...