Name: isolation and characterization of dendritic cells and macrophages from the mouse intestine
Uploaded: 2012-05-21T13:00:00
Description: Watch this Scientific Journal Video about Isolation and Characterization of Dendritic Cells and Macrophages from the Mouse Intestine at JoVE.com

我们感谢亚伦RAE（埃默里大学儿科和儿童医疗亚特兰大流核心），细胞分选。这项工作是由NIH资助AA01787001，从克隆氏病及溃疡性结肠炎美国基金会事业发展奖，和埃默里Egleston儿童研究中心种子批支持到顶级域名（TLD）

1。肠上皮细胞的解剖和离解制备的试剂和设备： <ul><li>温暖的Ca 2 + /镁2 +无PBS（CMF的PBS）的室温。 </li><li>温暖的Ca 2 + / Mg 2 +的免费的HBSS 5％胎牛血清（CMF方案的HBSS / FBS），和2mM EDTA室温。 </li><li>温暖的轨道摇床至37°C。 </li></ul> 注：步骤1.1至1.7，必须尽快进行，以尽量减少细胞死亡的程度，并实现最大的细胞?...

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<img src="/files/ftp_upload/4040/4040fig3.jpg" alt="图3"/> 图3。直接组织消化时间的影响因素，重要的细胞产量和表面抗原表达的优化。细胞产量和表面抗原表达，胶原酶的具体特点，组织切碎程度，以及存在或炎症的情况下，这可能会影响组织的完整性和细胞结构。长期组织消化，可能会导致细胞活力下降和表面抗原的表达，同时组织消化不足可能会导致在分析细胞缺乏。 ...

<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> 1×PBS的Ca 2 + -和Mg 2 +无</td><td></td><td></td><td></td></tr><tr><td>汉克的平衡盐溶液（HBSS中）与酚红</td><td> Fisher Scientific则</td><td> SH3001603 </td><td></td></tr><tr><td>碳酸氢钠</td><td>西格玛</td><td> S6014 </td><td></td></tr><tr><td>在0.85％的NaCl 1M肝素钠</td><td>龙沙</td><td> 17-737E </td><td></td></tr><tr><td>胎牛血清（FBS）， </td><td>亚特兰大生物制品</td><td> S11150H </td><td>热灭活</td></tr><tr><td> 0.5M EDTA的（pH值8.0） </td><td> cellgro </td><td> 46-034-CI </td><td></td></tr><tr><td> collagenasE型八</td><td>西格玛</td><td> C2139 </td><td></td></tr><tr><td> DNA酶I </td><td>罗氏公司</td><td> 14785000 </td><td>联合解决方案：100mg/ml </td></tr><tr><td> LIVE / DEAD可修复水族死细胞染色试剂盒，为405 nm激发</td><td> Invitrogen公司</td><td> L34957 </td><td>使用1:1000 </td></tr><tr><td> CD45 - ，PerCP单克隆抗体（30F11） </td><td>屋宇署</td><td> 557235 </td><td>使用1:100 </td></tr><tr><td> CD103-PE单克隆抗体（M290） </td><td>屋宇署</td><td> 557495 </td><td>使用1:100 </td></tr><tr><td> FcγRIII/ II抗体（2.4G2） </td><td>屋宇署</td><td> 553141 </td><td>在1:200的使用</td></tr><tr><td> CD11c的APC的单克隆抗体（N418） </td><td> eBioscience </td><td> 17-0114-82 </td><td>使用1:100 </td></tr><tr><td> MHC-Ⅱ（我-AB）的Alexa Fluor 700单克隆抗体</td><td> eBioscience </td><td> 56-5321-82 </td><td>使用1:100 </td></tr><tr><td>细胞CD11b-eFluor 450单克隆抗体（M1/70） </td><td> eBioscience </td><td> 48-0112-82 </td><td>在1:200的使用</td></tr><tr><td> F4/80-PE-Cy7单克隆抗体（BM8） </td><td> eBioscience </td><td> 25-4801-82 </td><td></td></tr><tr><td> CD11b的微球</td><td>美天旎生物技术</td><td> 130-049-601 </td><td></td></tr><tr><td> CD11c的微球</td><td>美天旎生物技术</td><td> 130-052-001 </td><td></td></tr><tr><td> 50 ml锥形管</td><td>屋宇署猎鹰</td><td> 352098 </td><td></td></tr><tr><td>单铁丝网过滤</td><td> Chefmate </td><td></td><td></td></tr><tr><td>小重量船</td><td> Fisher Scientific则</td><td> 08-732-116 </td><td></td></tr><tr><td> 100微米的细胞过滤器</td><td>屋宇署猎鹰</td><td> 352360 </td><td></td></tr><tr><td> 40微米的细胞过滤器</td><td>屋宇署猎鹰</td><td> 352340 </td><td></td></tr><tr><td> 5毫升聚苯乙烯圆底管</td><td>屋宇署猎鹰</td><td> 352235 </td><td>使用1:100 </td></tr><tr><td> MAXQ 4450台式轨道摇床</td><td> Thermo Scientific的</td><td></td><td></td></tr><tr><td>的LS磁珠列</td><td>美天旎生物技术</td><td> 130-042-401 </td><td></td></tr><tr><td> LSR的二</td><td>屋宇署</td><td></td><td></td></tr><tr><td> FACSAria二</td><td>屋宇署</td><td></td><td></td></tr></tbody></table>

isolation and characterization of dendritic cells and macrophages from the mouse intestine

肠道内居住的是参与对共生的植物和食物抗原在促进容忍，而随之而来的剩余准备安装的炎症反应，病原体侵入朝1,2先天免疫和适应性免疫细胞的独特群体。抗原提呈细胞，特别是区议会和巨噬细胞，发挥关键作用，在肠道免疫稳态维持通过自己的能力感和妥善应对微生物3-14。肠道DC和巨噬细胞的高效分离是在描述这些细胞的表型和功能的关键一步。虽然隔离肠道免疫细胞，包括区议会和巨噬细胞，许多有效的方法已6,10,15-24，许多依靠长期的消化时间，可能会产生负面影响细胞表面抗原的表达，细胞活力，和/或细胞产量。在这里，我们详细介绍快速分离的大量viabl的一种方法E，肠区议会和巨噬细胞。肠道DC和巨噬细胞的表型特征进行了直接染色孤立的肠细胞与特定的荧光标记单克隆抗体多色流式细胞仪分析。此外，高纯度的DC和巨噬细胞群体隔离功能的研究，利用CD11c和CD11b的磁激活细胞分选，细胞分选微珠。

Watch this Scientific Journal Video about Isolation and Characterization of Dendritic Cells and Macrophages from the Mouse Intestine at JoVE.com

Isolation and Characterization of Dendritic Cells and Macrophages from the Mouse Intestine

在这里，我们详细介绍了小鼠肠道树突状细胞（DCs）和巨噬细胞的快速分离方法。使用多色流式细胞仪分析细胞分选磁珠富集而被用来产生高纯度的功能研究人口肠道DC和巨噬细胞的表型特征。

从老鼠肠道树突状细胞和巨噬细胞的分离和表征

Within the intestine reside unique populations of innate and adaptive immune cells that are involved in promoting tolerance towards commensal flora and ...

Research

JoVE Journal

Immunology and Infection

Isolation of Mouse Lung Dendritic Cells

小鼠肺树突状细胞的分离

Lung dendritic cells (DC) play a fundamental role in sensing invading pathogens 1,2 as well as in the control of tolerogenic responses 3 in the ...

科研

免疫与感染

Isolation of Myeloid Dendritic Cells and Epithelial Cells from Human Thymus

从人类胸腺髓样树突状细胞和上皮细胞的分离

In this protocol we provide a method to isolate dendritic cells (DC) and epithelial cells (TEC) from the human thymus. DC and TEC are the major antigen ...

T Cells Capture Bacteria by Transinfection from Dendritic Cells

性T细胞Transinfection从树突状细胞捕捉细菌

Recently, we have shown, contrary to what is described, that CD4+ T cells, the paradigm of adaptive immune cells, capture bacteria from infected dendritic ...

Isolation and Flow Cytometric Analysis of Immune Cells from the Ischemic Mouse Brain

分离和流式细胞仪分析从缺血小鼠脑免疫细胞

Ischemic stroke initiates a robust inflammatory response that starts in the intravascular compartment and involves rapid activation of brain resident ...

Generation and Identification of GM-CSF Derived Alveolar-like Macrophages and Dendritic Cells From Mouse Bone Marrow

代和GM-CSF的鉴定源性肺泡状巨噬细胞和树突状细胞从小鼠骨髓

Macrophages and dendritic cells (DCs) are innate immune cells found in tissues and lymphoid organs that play a key role in the defense against pathogens. ...

Characterization of Human Monocyte-derived Dendritic Cells by Imaging Flow Cytometry: A Comparison between Two Monocyte Isolation Protocols

通过成像流式细胞仪人单核细胞来源的树突状细胞的特征：两个单核细胞分离方法的比较

Dendritic cells (DCs) are antigen presenting cells of the immune system that play a crucial role in lymphocyte responses, host defense mechanisms, and ...

Phenotypic Characterization of Macrophages from Rat Kidney by Flow Cytometry

通过流式细胞仪大鼠肾脏巨噬细胞的表型特征

There is increasing evidence suggesting the important role of inflammation and, subsequently, macrophages in the development and progression of renal ...

Fluorescence-activated Cell Sorting for Purification of Plasmacytoid Dendritic Cells from the Mouse Bone Marrow

荧光激活细胞分选从小鼠骨髓浆树突状细胞的分离纯化

Fluorescence-activated cell sorting (FACS) is a technique to purify specific cell populations based on phenotypes detected by flow cytometry. This method ...

Generation of Human Monocyte-derived Dendritic Cells from Whole Blood

从全血人单核细胞来源的树突状细胞的生成

Dendritic cells (DCs) recognize foreign structures of different pathogens, such as viruses, bacteria, and fungi, via a variety of pattern recognition ...

Isolation, Characterization, and Purification of Macrophages from Tissues Affected by Obesity-related Inflammation

受肥胖相关炎症组织的巨噬细胞的分离、鉴定和纯化

Obesity promotes a chronic inflammatory state that is largely mediated by tissue-resident macrophages as well as monocyte-derived macrophages. ...