Name: isolation of functional cardiac immune cells
Uploaded: 2011-12-05T12:00:00
Description: Watch this Scientific Journal Video about Isolation of Functional Cardiac Immune Cells at JoVE.com

这项工作是由一个UNCF默克研究生学研究论文奖学金（JLM），国家心脏，肺，血液研究所授予RO1 - HL - 62228（JSJ）和RO1的HL - 073990（JSJ）的支持。

1。汉克斯“平衡盐溶液的制备（HBSS） <ol><li> 1000毫升高压灭菌器1升玻璃烧杯，量筒，搅拌棒。 </li><li>重组蒸压1 L玻璃烧杯用双蒸水900毫升标记的HBSS粉末的解决方案。 </li><li> HEPES钠+盐3.4克的HBSS混合溶液和pH值调整到7.35-7.41之间的价值，无论是使用浓盐酸或氢氧化钠。 </li><li>一旦所需的pH值已达到并保持稳定，加5克牛血清白蛋白在室温下的解决方案，直到完全溶解白蛋白。 </li><li>添加的抗生?...

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S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Pharmacologic+inhibition+of+mast+cell+degranulation+prevents+left+ventricular+remodeling+induced+by+chronic+volume+overload+in+rats.">Pharmacologic inhibition of mast cell degranulation prevents left ventricular remodeling induced by chronic volume overload in rats.</a> Journal of Cardiac Failure. 11, 548-556 (2005).</li><li>Chancey, A. L., Brower, G. L., Janicki, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+mast+cell-mediated+activation+of+gelatinase+and+alteration+of+ventricular+diastolic+function.">Cardiac mast cell-mediated activation of gelatinase and alteration of ventricular diastolic function.</a> Am. J. Physiol. Heart. Circ. Physiol. 282, H2152-H2158 (2002).</li><li>Levick, S. P., Gardner, J. D., Holland, M., Hauer-Jensen, M., Janicki, J. S., Brower, G. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Protection+from+adverse+myocardial+remodeling+secondary+to+chronic+volume+overload+in+mast+cell+deficient+rats.">Protection from adverse myocardial remodeling secondary to chronic volume overload in mast cell deficient rats.</a> Journal of Molecular and Cellular Cardiology. 43, 56-61 (2008).</li><li>Levick, S. P., McLarty, J. L., Murray, D. B., Freeman, R. M., Carver, W. E., Brower, G. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cardiac+Mast+Cells+Mediate+Left+Ventricular+Fibrosis+in+the+Hypertensive+Rat.">Cardiac Mast Cells Mediate Left Ventricular Fibrosis in the Hypertensive Rat.</a> Hypertension. 53, 1041-1047 (2009).</li><li>Batista, M. L., Santos, R. V., Cunha, L. M., Mattos, K., Oliveira, E. M., Seelander, M. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+the+pro-inflammatory+cytokine+production+and+peritoneal+macrophage+function+in+rats+with+chronic+heart+failure.">Changes in the pro-inflammatory cytokine production and peritoneal macrophage function in rats with chronic heart failure.</a> Cytokine. 34, 284-290 (2006).</li><li>Galli, S. J., Tsai, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mast+Cells:+Versatile+regulator+of+inflammation,+tissue+remodeling,+host+defense+and+homeostasis.">Mast Cells: Versatile regulator of inflammation, tissue remodeling, host defense and homeostasis.</a> Journal of Dermatological Science. 49, 7-19 (2008).</li><li>Yndestad, A., Holm, A. M., Muller, F., Simonsen, S., Froland, S. 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Physiol. 297, H1462-H1468 (2009).</li><li>Forman, M. F., Brower, G. L., Janicki, J. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Spontaneous+histamine+secretion+during+isolation+of+rat+cardiac+mast+cells.">Spontaneous histamine secretion during isolation of rat cardiac mast cells.</a> Inflammation Research. 53, 453-457 (2004).</li><li>Levick, S. P., Murray, D. B., Janicki, J. S., Brower, G. 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这种隔离心脏免疫细胞的方法，使无不需要激活，通常发生在传统的酶消化法分离技术，分析其功能，表型，成熟。这种技术获得的平均细胞总数约400000元大鼠心脏，差这些细胞的染色和流式细胞仪分析表明（〜70％），巨噬细胞（〜12％）和肥大细胞，T淋巴细胞混合的人口使用了这种技术。（〜12％）。13，14因此，心脏的免疫细胞群的进一步分离纯化方法是必要的。隔离的免疫细胞，可...

<table border="1"><tbody><tr><td>试剂和设备名称</td><td>公司</td><td>目录编号</td><td>评论</td></tr><tr><td>汉克斯“平衡盐</td><td>西格玛</td><td> H4891 </td><td>一瓶1 L汉克斯&#39;平衡盐 解决方案。重组汉克斯&#39;平衡盐 解决方案作为一个过程的一部分。 </td></tr><tr><td>抗生素的Antimyotic </td><td> Gibco公司</td><td> 15240-062 </td><td></td></tr><tr><td> HEPES </td><td>西格玛</td><td> H3784 </td><td></td></tr><tr><td>牛血清白蛋白</td><td>西格玛</td><td> A3912 </td><td></td></tr><tr><td> 250毫升Nalogene滤瓶</td><td> Thermoscientific </td><td> 157-0020 </td><td></td></tr><tr><td> 12 N盐酸</td><td> Fisher Scientific则</td><td>中科院7647-01-0 </td><td>对于6个N，添加盐酸等量去离子水。 </td></tr><tr><td> 10 N的氢氧化钠</td><td> Fisher Scientific则</td><td> SC267 </td><td> 5 N，添加去离子水相等体积的NaOH。 </td></tr><tr><td>双蒸水</td><td></td><td></td><td></td></tr></tbody></table>具体设备表： <table border="1"><tbody><tr><td>试剂和设备名称</td><td>公司</td><td>目录编号</td><td>评论</td></tr><tr><td> - 3 - O缝合丝绸编织</td><td>爱惜康产品</td><td> SA64H </td><td></td></tr><tr><td>小动物呼吸机</td><td> CWE公司</td><td> SAR - 830 </td><td>设置呼吸频率每分钟40.5次呼吸。 </td></tr><tr><td>斜角18号针头</td><td> BD公司</td><td> 305199 </td><td>修剪其原始长度的1 / 2至3 / 4使用气管套管针。 </td></tr><tr><td>聚四氟乙烯提示24计巴克斯特快速导管导管（带针头删除） </td><td>巴克斯特</td><td> 2N111 </td><td>聚四氟乙烯提示长度减少1 / 2至3 / 4。 </td></tr><tr><td> 10毫升注射器</td><td> Fisher Scientific则</td><td> 14 - 823 - 16B </td><td></td></tr><tr><td> 15毫升猎鹰管</td><td> BD公司</td><td> 352097 </td><td></td></tr><tr><td> Sorvall RT6000B离心机</td><td> Fisher Scientific则</td><td> 75-004-377 </td><td>记住预冷，并保持在4 ° C。 </td></tr><tr><td> Hyclone公司</td><td> Thermoscientific </td><td> S300030.02 </td><td></td></tr><tr><td>血球</td><td> Hausser科学</td><td> HS - LB - 1483 </td><td></td></tr></tbody></table>

isolation of functional cardiac immune cells

心脏免疫细胞获得利益的角色，他们在许多心脏疾病的病理重塑发挥1-5这些免疫细胞，其中包括肥大细胞，T细胞和巨噬细胞;存储和释放的多种生物活性介质，包括细胞因子和蛋白酶（如胰蛋白酶）。6-8这些调解员已被证明是通过激活基质金属蛋白酶或胶原蛋白的积累引起心脏成纤维细胞的功能，通过调节细胞外基质代谢的关键球员。9-11然而，隔离心脏免疫细胞技术一直是个问题，因为他们使用细菌胶原酶消化心肌组织。这种技术的原因，从而激活免疫细胞功能的损失。例如，心脏肥大细胞变得大大减少响应的化合物，导致脱粒， 因此 ，我们开发出一种技术，允许对T他隔离心脏功能的免疫细胞，这将导致更好地了解了这些细胞在心脏疾病中的作用。 13，14 此方法需要一个熟悉大鼠剑突，腋下和镰状韧带的解剖位置，对心脏和心包。这些标志性建筑是重要的，以提高手术成功和心脏的免疫细胞，以确保较高的产量。这些孤立的心脏，免疫细胞可以用于表征功能，表型，成熟，并与其他心肌细胞共培养实验，以获得一个更好地了解它们之间的相互作用。

Watch this Scientific Journal Video about Isolation of Functional Cardiac Immune Cells at JoVE.com

Isolation of Functional Cardiac Immune Cells

这种隔离的心脏功能的免疫细胞的方法提供了一种替代传统的方法胶原酶消化，从而导致不​​必要的免疫细胞活化，从而导致了这些细胞的减少响应。我们隔离的方法，产生心脏功能的免疫细胞，避免与酶消化相关的问题。

心脏功能免疫细胞的分离

Cardiac immune cells are gaining interest for the roles they play in the pathological remodeling in many cardiac diseases.1-5  These immune cells, which ...

Research

JoVE Journal

Immunology and Infection

Measuring Calpain Activity in Fixed and Living Cells by Flow Cytometry

在固定和活细胞，流式细胞仪测量钙蛋白酶活性

Calpains are ubiquitous intracellular, calcium-sensitive, neutral cysteine proteases 1. Calpains play crucial roles in many physiological processes, ...

科研

免疫与感染

Seven Steps to Stellate Cells

星状细胞的七个步骤

Hepatic stellate cells are liver-resident  cells of star-like morphology and are located in the space of Disse between  liver sinusoidal endothelial cells ...

Intravital Imaging of the Mouse Thymus using 2-Photon Microscopy

活体成像的鼠标使用2 - 光子显微镜的胸腺

Two-photon Microscopy (TPM) provides image acquisition in deep areas inside tissues and organs. In combination with the development of new stereotactic ...

Differentiating Functional Roles of Gene Expression from Immune and Non-immune Cells in Mouse Colitis by Bone Marrow Transplantation

区分功能基因表达的作用，通过骨髓移植免疫和非免疫细胞在小鼠结肠炎

To understand the role of a gene in the development of colitis, we compared the responses of wild-type mice and gene-of-interest deficient knockout mice ...

Flow Cytometric Isolation of Primary Murine Type II Alveolar Epithelial Cells for Functional and Molecular Studies

主要鼠Ⅱ型肺泡上皮细胞的功能及分子生物学研究的流式细胞仪分离

Throughout  the last years, the contribution of alveolar type II epithelial cells (AECII)  to various aspects of immune regulation in the lung has been ...

Isolation of Adipose Tissue Immune Cells

脂肪组织免疫细胞的分离

The discovery of increased macrophage infiltration in  the adipose tissue (AT) of obese rodents and humans has led to an  intensification of interest in ...

Detecting Glycogen in Peripheral Blood Mononuclear Cells with Periodic Acid Schiff Staining

与碘酸雪夫染色检测糖原的外周血单个核细胞

Periodic acid Schiff (PAS) staining is an immunohistochemical technique used on muscle biopsies and as a diagnostic tool for blood samples. ...

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 ...

Isolation, Characterization and Functional Examination of the Gingival Immune Cell Network

分离，鉴定和牙龈免疫细胞网络的功能性考试

Immune cell networks in tissues play a vital role in mediating local immunity and maintaining tissue homeostasis, yet little is known of the resident ...

Isolation and Characterization of Neutrophil-derived Microparticles for Functional Studies

中性粒细胞衍生微粒的分离与鉴定功能研究

Polymorphonuclear neutrophil-derived microparticles (PMN)-MPs) are lipid bilayer, spherical microvesicles with sizes ranging from 50&#8211;1,000 nm in ...