Name: depletion and reconstitution of macrophages in mice
Uploaded: 2012-08-01T13:00:00
Description: Watch this Scientific Journal Video about Depletion and Reconstitution of Macrophages in Mice at JoVE.com

这项工作得到了1“的研究资助津贴”的克隆氏病的和结肠炎加拿大基金会的LMS，谁是1​​消化科/加拿大协会健康研究/克罗恩的小号和结肠炎加拿大新研究者奖基金会加拿大协会的支持。

1。使用含有氯磷酸二钠，脂质体的消耗巨噬细胞<ol><li>脂质体储存在4°C注射前两个小时，从冰箱中取出氯膦酸二钠含脂质体，PBS含脂质体，或无菌PBS（喷射控制），使他们能够适应新环境室温（18℃）。 </li><li>反转管含脂质体的8-10倍，以确保均匀分布，然后装入1 ml注射器200μL。将26号针头注射器顶端。 </li><li>用你的左手，浮渣鼠标，略低于持有足够的皮肤以固定其头部和四肢的耳朵。 </li><li>...

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	<li>Gordon, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Alternative+activation+of+macrophages.">Alternative activation of macrophages.</a> Nat. Rev. Immunol. 3, 23-35 (2003).</li><li>Rescigno, M., Lopatin, U., Chieppa, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Interactions+among+dendritic+cells,+macrophages,+and+epithelial+cells+in+the+gut:+implications+for+immune+tolerance.">Interactions among dendritic cells, macrophages, and epithelial cells in the gut: implications for immune tolerance.</a> Curr. Opin. Immunol. 20, 669-675 (2008).</li><li>Heinsbroek, S. 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B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=SHIP-deficient,+alternatively+activated+macrophages+protect+mice+during+DSS-induced+colitis.">SHIP-deficient, alternatively activated macrophages protect mice during DSS-induced colitis.</a> J. Leukoc. Biol. 90, 483-492 (2011).</li><li>Qualls, J. E., Kaplan, A. M., van Rooijen, N., Cohen, D. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Suppression+of+experimental+colitis+by+intestinal+mononuclear+phagocytes.">Suppression of experimental colitis by intestinal mononuclear phagocytes.</a> J. Leukoc. 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巨噬细胞的吞噬细胞在免疫系统中发挥重要作用。他们是负责发起的先天免疫反应和指挥后天免疫反应。通常情况下，经典激活巨噬细胞被激活IFNγ的或LPS，并有责任为消除病原体和安装的炎症反应1。相反，IL-4和IL-13激活或者激活巨噬细胞和炎症的决议过程中发挥在碎片中的作用，清除和组织重塑。专门的巨噬细胞在肠道保留其杀菌活性，但不装入一个亲炎症反应2。这种有益菌?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td>氯膦酸二钠，含脂质体</td><td> <a href="http://Clodronateliposomes.org" target="_blank">Clodronateliposomes.org</a> </td><td></td></tr><tr><td> PBS含脂质体</td><td> <a href="http://Clodronateliposomes.org" target="_blank">Clodronateliposomes.org</a> </td><td></td></tr><tr><td>无菌PBS pH7.4的</td><td> Invitrogen公司</td><td> 14190 </td></tr><tr><td> IMDM培养</td><td> Invitrogen公司</td><td> 12440 </td></tr><tr><td>胎牛血清（FCS）的</td><td> Invitrogen公司</td><td> 12483 </td></tr><tr><td>醋酸</td><td> BDH问题Aristar </td><td> BDH3092-500MLP </td></tr><tr><td> monothioglycerol胺酶（MTG） </td><td>西格玛</td><td> 96-275 </td></tr><tr><td> RECOMBinant小鼠MCSF </td><td>干细胞技术</td><td> 02951 </td></tr><tr><td>重组鼠GM-CSF的</td><td>干细胞技术</td><td> 02935 </td></tr><tr><td>重组小鼠IL-3 </td><td>干细胞技术</td><td> 02903 </td></tr><tr><td>重组小鼠γ干扰素</td><td>干细胞技术</td><td> 02746 </td></tr><tr><td>重组小鼠IL-4 </td><td>干细胞技术</td><td> 02714 </td></tr><tr><td>细胞分离缓冲器</td><td> Invitrogen公司</td><td> 13150-016 </td></tr><tr><td>大鼠anti-F4/80抗体</td><td> ABD Serotec公司</td><td> MCA497GA </td></tr><tr><td>鼠抗ArgI抗体</td><td> BD转导实验室</td><td> 610708 </td></tr></tbody></table> 表1。在本协议中使用的特定试剂。

depletion and reconstitution of macrophages in mice

巨噬细胞是重要的球员，在传染病的挑战或损伤的先天免疫反应，发起的先天免疫反应和指挥后天免疫反应。巨噬细胞功能障碍可能导致无法安装适当的免疫反应，因此，已在许多疾病的进程，包括炎症性肠病牵连。巨噬细胞显示极化表型大致分为两类。经典活化的巨噬细胞，与干扰素γ或LPS刺激激活，发挥了至关重要的作用，在细菌的挑战，而另外激活的巨噬细胞，IL-4和IL-13激活，参与清除杂物和组织重塑和已在该决议牵连阶段的炎症。在体内的炎症反应，巨噬细胞的免疫细胞浸润的复杂混合物中发现，并可能加剧或resolvin参加Ğ炎症。巨噬细胞的作用，在整个动物模型的定义在原地 ，这是必要的检查消耗从复杂的环境中的巨噬细胞的影响。极化的巨噬细胞表型定义问原位巨噬细胞表型的作用的问题，可以导出体外 ，从骨髓穿刺，并加回或没有事先枯竭的巨噬细胞，小鼠。在这里提出的协议中含有氯膦酸二钠，脂质体，与PBS注射控件，用来消耗结肠巨噬细胞在葡聚糖硫​​酸钠（DSS）诱导小鼠结肠炎。此外，巨噬细胞的极化，推导出体外，并转移到小鼠静脉注射。这种方法的一个需要注意的是，氯膦酸二钠含脂质体耗尽所有专业的吞噬细胞，包括树突状细胞和巨噬细胞，所以，以确保枯竭所观察到的效果是巨噬细胞特异性，重组ØF的巨噬细胞过继转移表型是必要的。全身巨噬细胞耗竭小鼠也可以通过回交小鼠实现到细胞CD11b-DTR背景，这是一个很好的补充办法。含有氯膦酸二钠脂质体介导的枯竭的优势是，它不参与回交小鼠需要的时间和费用，它可以在小鼠小鼠（C57BL / 6小鼠，BALB / c小鼠，或混合背景的背景下，无论使用）。

Watch this Scientific Journal Video about Depletion and Reconstitution of Macrophages in Mice at JoVE.com

Depletion and Reconstitution of Macrophages in Mice

巨噬细胞发挥核心作用的动态平衡，并在许多组织的病理。这里介绍的协议描述消耗巨噬细胞的方法<em&gt;在体内</em&gt;，来自偏光巨噬细胞，骨髓穿刺，继转移到小鼠巨噬细胞。这些技术允许极化的巨噬细胞在健康和疾病中发挥的作用的决心。

在小鼠体内的巨噬细胞的枯竭和重建

Macrophages are critical players in the innate  immune response to infectious challenge or injury, initiating the innate immune  response and directing ...

Research

JoVE Journal

Immunology and Infection

Depletion of Specific Cell Populations by Complement Depletion

补消耗的特定细胞群的枯竭

The purification of immune cell populations is often required in order to study their unique functions.  In particular, molecular approaches such as ...

科研

免疫与感染

Study of Phagolysosome Biogenesis in Live Macrophages

在现场研究巨噬细胞吞噬溶酶体生源

Phagocytic cells play a major role in the innate immune system by removing and eliminating invading microorganisms in their phagosomes. Phagosome ...

Use of Shigella flexneri to Study Autophagy-Cytoskeleton Interactions

Use of Shigella flexneri to Study Autophagy-Cytoskeleton Interactions

使用<em&gt;福氏痢疾杆菌</em&gt;研究自噬，细胞骨架相互作用

Shigella flexneri is an intracellular pathogen that can escape from phagosomes to reach the cytosol, and polymerize the host actin cytoskeleton to promote ...

Proteomic Profiling of Macrophages by 2D Electrophoresis

巨噬细胞通过二维电泳蛋白组学分析

The goal of the two-dimensional (2D) electrophoresis protocol described here is to show how to analyse the phenotype of human cultured macrophages. The ...

Macrophage Cholesterol Depletion and Its Effect on the Phagocytosis of Cryptococcus neoformans

Macrophage Cholesterol Depletion and Its Effect on the Phagocytosis of Cryptococcus neoformans

巨噬细胞胆固醇消耗及其对的吞噬作用<em&gt;新型隐球菌</em

Cryptococcosis is a life-threatening infection caused by pathogenic fungi of the genus Cryptococcus. Infection occurs upon inhalation of spores, which are ...

Quantification of Cytosolic vs. Vacuolar Salmonella in Primary Macrophages by Differential Permeabilization

Quantification of Cytosolic vs. Vacuolar Salmonella in Primary Macrophages by Differential Permeabilization

胞浆的定量与液泡<em&gt;沙门氏菌</em&gt;在原发性巨噬细胞差透

Intracellular bacterial pathogens can replicate in the cytosol or in specialized pathogen-containing vacuoles (PCVs). To reach the cytosol, bacteria like ...

Isolation of Salmonella typhimurium-containing Phagosomes from Macrophages

Isolation of Salmonella typhimurium-containing Phagosomes from Macrophages

巨噬细胞中含有吞噬的鼠伤寒沙门氏菌的分离

Salmonella typhimurium is a facultative intracellular bacterium that causes gastroenteritis in humans. After invasion of the lamina propria, S. ...

Isolation and In Vitro Culture of Murine and Human Alveolar Macrophages

Isolation and In Vitro Culture of Murine and Human Alveolar Macrophages

小鼠与人肺泡巨噬细胞的分离和体外培养

Alveolar macrophages are terminally differentiated, lung-resident macrophages of prenatal origin. Alveolar macrophages are unique in their long life and ...

Flow Cytometric Measurement Of ROS Production In Macrophages In Response To Fc&#947;R Cross-linking

Flow Cytometric Measurement Of ROS Production In Macrophages In Response To FcÃƒÅ½Ã‚Â³R Cross-linking

巨噬细胞 ros 产生的流式细胞仪测量对 fcγr 交联的响应

The oxidative or respiratory burst is used to describe the rapid consumption of oxygen and generation of reactive oxygen species (ROS) by phagocytes in ...

Quantification of Monocyte Chemotactic Activity In Vivo and Characterization of Blood Monocyte Derived Macrophages

单核细胞化学策略活性在Vivo的定量和血液单核细胞衍生巨噬细胞的特征

Tissue homeostasis and repair are critically dependent on the recruitment of monocyte-derived macrophages. Both under- and over-recruitment of ...