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

यह काम एक &#39;रिसर्च सहायता में अनुदान&#39; है Crohn है और कनाडा के बृहदांत्रशोथ फाउंडेशन से LMS, जो / गैस्ट्रोएंटरोलॉजी कैनेडियन स्वास्थ्य / अनुसंधान Crohn है &#39;एस के एसोसिएशन और कनाडा नई अन्वेषक पुरस्कार के बृहदांत्रशोथ फाउंडेशन के एक कनाडा के एसोसिएशन द्वारा समर्थित है के द्वारा समर्थित किया गया था.

1. घट Clodronate - युक्त Liposomes का उपयोग मैक्रोफेज <ol><li> Liposomes 4 डिग्री सेल्सियस पर जमा हो जाती है इंजेक्शन के लिए दो घंटे पहले, रेफ्रिजरेटर से clodronate युक्त liposomes, पीबीएस युक्त liposomes, या बाँझ पीबीएस (इंजेक्शन नियंत्रण) को हटाने...

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मैक्रोफेज phagocytic कोशिकाओं है कि प्रतिरक्षा प्रणाली में एक महत्वपूर्ण भूमिका निभाते हैं. वे सहज प्रतिरक्षा प्रतिक्रिया शुरू करने और अर्जित प्रतिरक्षा प्रतिक्रिया निर्देशन के लिए जिम्मेदार हैं. आमतौर ?...

<table border="1"><tbody><tr><td> अभिकर्मक का नाम </td><td> कंपनी </td><td> सूचीपत्र संख्या </td></tr><tr><td> Clodronate - युक्त liposomes </td><td> <a href="http://Clodronateliposomes.org" target="_blank">Clodronateliposomes.org</a> </td><td></td></tr><tr><td> पीबीएस - युक्त liposomes </td><td> <a href="http://Clodronateliposomes.org" target="_blank">Clodronateliposomes.org</a> </td><td></td></tr><tr><td> बाँझ पीबीएस pH7.4 </td><td> Invitrogen </td><td> 14,190 </td></tr><tr><td> IMDM </td><td> Invitrogen </td><td> +१२४४० </td></tr><tr><td> भ्रूण बछड़ा सीरम (एफसीएस) </td><td> Invitrogen </td><td> 12,483 </td></tr><tr><td> एसिटिक एसिड </td><td> BDH Aristar </td><td> BDH3092 - 500MLP </td></tr><tr><td> Monothioglycerol (MTG) </td><td> सिग्मा </td><td> 96-275 </td></tr><tr><td> RECOMBinant murine MCSF </td><td> Stemcell टेक्नोलॉजीज </td><td> 02,951 </td></tr><tr><td> पुनः संयोजक जीएम-CSF murine </td><td> Stemcell टेक्नोलॉजीज </td><td> 02,935 </td></tr><tr><td> पुनः संयोजक murine आईएल -3 </td><td> Stemcell टेक्नोलॉजीज </td><td> 02,903 </td></tr><tr><td> पुनः संयोजक murine IFNγ </td><td> Stemcell टेक्नोलॉजीज </td><td> 02,746 </td></tr><tr><td> पुनः संयोजक murine आईएल 4 </td><td> Stemcell टेक्नोलॉजीज </td><td> 02,714 </td></tr><tr><td> सेल हदबंदी बफर </td><td> Invitrogen </td><td> 13150-016 </td></tr><tr><td> Anti-F4/80 एंटीबॉडी चूहे </td><td> अब्द Supplier: </td><td> MCA497GA </td></tr><tr><td> माउस एंटीबॉडी विरोधी - ARGI </td><td> बी.डी. पारगमन लेबोरेटरीज </td><td> 610,708 </td></tr></tbody></table> तालिका 1 विशिष्ट अभिकर्मकों इस प्रोटोकॉल में प्रयोग किया जाता है.

depletion and reconstitution of macrophages in mice

Macrophages are critical players in the innate immune response to infectious challenge or injury, initiating the innate immune response and directing the acquired immune response. Macrophage dysfunction can lead to an inability to mount an appropriate immune response and as such, has been implicated in many disease processes, including inflammatory bowel diseases. Macrophages display polarized phenotypes that are broadly divided into two categories. Classically activated macrophages, activated by stimulation with IFN&gamma; or LPS, play an essential role in response to bacterial challenge whereas alternatively activated macrophages, activated by IL-4 or IL-13, participate in debris scavenging and tissue remodeling and have been implicated in the resolution phase of inflammation. During an inflammatory response in vivo, macrophages are found amid a complex mixture of infiltrating immune cells and may participate by exacerbating or resolving inflammation. To define the role of macrophages in situ in a whole animal model, it is necessary to examine the effect of depleting macrophages from the complex environment. To ask questions about the role of macrophage phenotype in situ, phenotypically defined polarized macrophages can be derived ex vivo, from bone marrow aspirates and added back to mice, with or without prior depletion of macrophages. In the protocol presented here clodronate-containing liposomes, versus PBS injected controls, were used to deplete colonic macrophages during dextran sodium sulfate (DSS)-induced colitis in mice. In addition, polarized macrophages were derived ex vivo and transferred to mice by intravenous injection. A caveat to this approach is that clodronate-containing liposomes deplete all professional phagocytes, including both dendritic cells and macrophages so to ensure the effect observed by depletion is macrophage-specific, reconstitution of phenotype by adoptive transfer of macrophages is necessary. Systemic macrophage depletion in mice can also be achieved by backcrossing mice onto a CD11b-DTR background, which is an excellent complementary approach. The advantage of clodronate-containing liposome-mediated depletion is that it does not require the time and expense involved in backcrossing mice and it can be used in mice regardless of the background of the mice (C57BL/6, BALB/c, or mixed background).

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

Depletion and Reconstitution of Macrophages in Mice

मैक्रोफेज homeostasis में एक केंद्रीय भूमिका निभाते हैं और कई ऊतकों में विकृति. यहाँ प्रस्तुत प्रोटोकॉल घट मैक्रोफेज के लिए तरीकों का वर्णन<em&gt; Vivo में</em, अस्थि मज्जा रेस्पायरेट्रस से polarized मैक्रोफेज पाने, और adoptively चूहों में मैक्रोफेज स्थानांतरित. इन तकनीकों में भूमिका का दृढ़ संकल्प है कि polarized मैक्रोफेज स्वास्थ्य और रोग में खेलने की अनुमति देते हैं.

और चूहे में मैक्रोफेज की कमी पुनर्गठन

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

विशिष्ट सेल आबादी के पूरक रिक्तीकरण द्वारा रिक्तीकरण

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

Study of Phagolysosome Biogenesis in Live Macrophages

लाइव मैक्रोफेज में Phagolysosome biogenesis का अध्ययन

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

का प्रयोग करें<em&gt; शिगेला flexneri</em&gt; भोजी-cytoskeleton बातचीत का अध्ययन

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

2 डी वैद्युतकणसंचलन द्वारा मैक्रोफेज की प्रोटिओमिक रूपरेखा

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

बृहतभक्षककोशिका कोलेस्ट्रॉल कमी और के phagocytosis पर उसके प्रभाव<em&gt; क्रिप्टोकोकस neoformans</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

Vacuolar बनाम साइटोसोलिक की मात्रा का ठहराव<em&gt; साल्मोनेला</em&gt; विभेदक Permeabilization द्वारा प्राथमिक मैक्रोफेज में

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

अलगाव साल्मोनेला typhimurium-मैक्रोफेज से युक्त Phagosomes का

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

अलगाव और इन विट्रो संस्कृति Murine और मानव वायुकोशीय मैक्रोफेज

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

fcγr पार जोड़ने के जवाब में मैक्रोफेज में ROS उत्पादन के प्रवाह cytometric माप

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

विवो में मोनोसाइट केमोटेक्टिक गतिविधि का परिमाणीकरण और रक्त मोनोसाइट व्युत्पन्न मैक्रोफेज की विशेषता

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