Name: isolation, purification and labeling of mouse bone marrow neutrophils for functional studies and adoptive transfer experiments
Uploaded: 2013-07-10T13:00:00
Description: Watch this Scientific Journal Video about Isolation, Purification and Labeling of Mouse Bone Marrow Neutrophils for Functional Studies and Adoptive Transfer Experiments at JoVE.com

这项工作是支持该司院内研究的美国国家过敏和传染病研究所（NIAID），国立卫生研究院（NIH），美国。 所有小鼠均保持在美国协会认可的实验室动物护理动物设施在国家过敏和传染病研究所（NIAID）的评审，并根据实验室动物护理和使用指南中列出的程序安置在的主持下，由动物护理和使用委员会NIAID批准的协议。

1。小鼠骨髓细胞的分离<ol><li>安乐死小鼠使用该机构的动物保健委员会批准的协议，和动物表面喷用70％的乙醇。 </li><li>在中腹部的皮肤做一个切口，去除皮肤的鼠标从远端部分包括下肢皮肤覆盖。 </li><li>由下肢用剪刀切断肌肉和小心，同时避免破坏股骨头从髋关节脱位的髋臼。 </li><li>取出剩余使用手术刀和剪刀从股骨和胫骨的肌肉和上面的膝关节多加小心，不破骨端部从胫骨股骨分离。放?...

<ol>
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S., Souto-Carneiro, M. M., Fonseca, J. E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neutrophils+in+rheumatoid+arthritis:+More+than+simple+final+effectors.">Neutrophils in rheumatoid arthritis: More than simple final effectors.</a> Autoimmun. Rev. 9 (8), 531-535 (2010).</li><li>Denny, M. F., Yalavarthi, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+distinct+subset+of+proinflammatory+neutrophils+isolated+from+patients+with+systemic+lupus+erythematosus+induces+vascular+damage+and+synthesizes+type+I+IFNs.">A distinct subset of proinflammatory neutrophils isolated from patients with systemic lupus erythematosus induces vascular damage and synthesizes type I IFNs.</a> J. Immunol. 184 (6), 3284-3297 (2010).</li><li>Kim, N. D., Chou, R. C., Seung, E., Tager, A. M., Luster, A. 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F., Verzaal, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neutrophils+are+indispensable+for+hematopoietic+stem+cell+mobilization+induced+by+interleukin-8+in+mice.">Neutrophils are indispensable for hematopoietic stem cell mobilization induced by interleukin-8 in mice.</a> Proc. Natl. Acad. Sci. U.S.A. 99 (9), 6228-6233 (2002).</li><li>Eash, K. J., Greenbaum, A. M., Gopalan, P. K., Link, D. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=CXCR2+and+CXCR4+antagonistically+regulate+neutrophil+trafficking+from+murine+bone+marrow.">CXCR2 and CXCR4 antagonistically regulate neutrophil trafficking from murine bone marrow.</a> J. Clin. Invest. 120 (7), 2423-2431 (2010).</li><li>Lionakis, M. S., Lim, J. K., Lee, C. C., Murphy, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Organ-specific+innate+immune+responses+in+a+mouse+model+of+invasive+candidiasis.">Organ-specific innate immune responses in a mouse model of invasive candidiasis.</a> J. Innate. Immun. 3 (2), 180-199 (2011).</li><li>Boxio, R., Bossenmeyer-Pouri&#233;, C., Steinckwich, N., Dournon, C., N&#252;sse, O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+bone+marrow+contains+large+numbers+of+functionally+competent+neutrophils.">Mouse bone marrow contains large numbers of functionally competent neutrophils.</a> J. Leukoc. Biol. 75 (4), 604-611 (2004).</li><li>Kim, N. D., Chou, R. C., Seung, E., Tager, A. M., Luster, A. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+unique+requirement+for+the+leukotriene+B4+receptor+BLT1+for+neutrophil+recruitment+in+inflammatory+arthritis.">A unique requirement for the leukotriene B4 receptor BLT1 for neutrophil recruitment in inflammatory arthritis.</a> J. Exp. Med. 203 (4), 829-835 (2006).</li><li>Gunzer, M., Weishaupt, C., Planelles, L., Grabbe, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Two-step+negative+enrichment+of+CD4++and+CD8++T+cells+from+murine+spleen+via+nylon+wool+adherence+and+an+optimized+antibody+cocktail.">Two-step negative enrichment of CD4+ and CD8+ T cells from murine spleen via nylon wool adherence and an optimized antibody cocktail.</a> J. Immunol. Methods. 258 (1-2), 55-63 (2001).</li><li>Tosello Boari, J., Amezcua Vesely, M. C., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=IL-17RA+signaling+reduces+inflammation+and+mortality+during+Trypanosoma+cruzi+infection+by+recruiting+suppressive+IL-10-producing+neutrophils.">IL-17RA signaling reduces inflammation and mortality during Trypanosoma cruzi infection by recruiting suppressive IL-10-producing neutrophils.</a> PLoS Pathog. 8 (4), e1002658 (2012).</li><li>Hattori, H., Subramanian, K. K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small-molecule+screen+identifies+reactive+oxygen+species+as+key+regulators+of+neutrophil+chemotaxis.">Small-molecule screen identifies reactive oxygen species as key regulators of neutrophil chemotaxis.</a> Proc. Natl. Acad. Sci. U.S.A. 107 (8), 3546-3351 (2010).</li><li>Wan, W., Lionakis, M. S., Liu, Q., Roff&#234;, E., Murphy, P. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genetic+Deletion+of+Chemokine+Receptor+Ccr7+Exacerbates+Atherogenesis+in+ApoE-deficient+Mice.">Genetic Deletion of Chemokine Receptor Ccr7 Exacerbates Atherogenesis in ApoE-deficient Mice.</a> Cardiovasc. Res. 97 (3), 580-588 (2013).</li><li>Ueda, Y., Kondo, M., Kelsoe, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inflammation+and+the+reciprocal+production+of+granulocytes+and+lymphocytes+in+bone+marrow.">Inflammation and the reciprocal production of granulocytes and lymphocytes in bone marrow.</a> J. Exp. Med. 201 (11), 1771-1780 (2005).</li><li>Berkow, R. L., Dodson, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Purification+and+functional+evaluation+of+mature+neutrophils+from+human+bone+marrow.">Purification and functional evaluation of mature neutrophils from human bone marrow.</a> Blood. 68 (4), 853-860 (1986).</li></ol>

在这里，我们提出了一个可靠，简单，快速，经济的协议，用于隔离大量嗜中性粒细胞从骨髓的小鼠具有纯度高，使用密度梯度离心法的可行性。 〜6-12×10 6中性粒细胞正确执行此协议，可以从收回未受感染的小鼠，从小鼠在感染后6和多达〜30-40×10 6中性粒细胞可能被孤立。分离出的嗜中性粒细胞是80-95％的纯&gt; 95％是可行的。 骨髓是一种合适的水库...

嗜中性粒细胞是人类最丰富的白细胞。他们是主要的先天免疫系统和抵御入侵的微生物的第一线作为细胞成分。获得性中性粒细胞减少症和原发性免疫缺陷的患者，影响中性粒细胞数量和/或功能的发展危及生命的侵袭性细菌和真菌感染，这些细胞在宿主防御1突出的重要性。在一场精心策划的先天免疫反应，从而导致分泌趋化因子产生趋化梯度能够招募中性粒细胞从血液中进入发炎组织2诱导其同源的模式识别受体的结果入侵的病原体在感染部位的免疫识别。中性粒细胞进入感染部位后，他们成为激活，导致细胞因子和趋化因子的生产，病原体吸收，并通过氧化和非氧化我杀chanisms 3。除了 ​​大家公认的先天免疫的作用，中性粒细胞也被最近表现出作为有效的适应性免疫反应的发起人，扮演着重要的角色。另一方面，从它们的免疫保护作用外，中性粒细胞也可介导组织损伤和由于过度积累和/或激活在炎症部位 ​​的免疫病理学，如图中的各种感染和自身免疫性疾病5-7。 尽管中性粒细胞在安装有效的先天免疫反应和一些传染性疾病和炎症性疾病，技术上的困难，处理这些细胞，并缺乏可靠的实验协议的多效性效应功能不可或缺的作用受阻的研究，在过去的几十年里，中性粒细胞。因此，应进一步促进使用可重复的检测，隔离中性粒细胞中性粒细胞介导的​​immunolo研究gical功能体外和体内 。迄今为止，已经描述的几种方法已用于分离，如密度梯度离心的人血和小鼠血液或骨髓8,9，正极或负极的从小鼠血液或骨髓10,11中的嗜中性粒细胞的免疫磁性富集的嗜中性粒细胞，和收获巯基乙酸或其他抗炎剂12后，腹腔注射的小鼠的腹腔内中性粒细胞。虽然可以很容易地从人血中分离出大量中性粒细胞，这种方法是最理想的小鼠由于数量有限小鼠血液，排除隔离足够的中性粒细胞功能的研究或过继转移实验13。此外，虽然产量的巯基乙酸盐诱导的腹膜腔中的细胞相比，小鼠血嗜中性粒细胞在炎症腹腔灌洗，纯度之间变化60-90％，分离出的嗜中性粒细胞，表现出一个激活的表型。因此，收集的细胞用这种方法只能用于执行功能的激活的研究，但并不代表未受刺激的嗜中性粒细胞，小鼠腹膜腔有中性粒细胞数，在稳定状态12。相反，骨髓是一个方便的水库收获大量的任何未受刺激或激活的中性粒细胞11,14，它可以被用于下游功能性研究，如吞噬作用，杀害和脱颗粒，或过继转移到受体小鼠。 在这里，我们介绍一个简单而快速的协议（约2小时），它提供了一个高收益率（〜6-12×10 6中性粒细胞/未受感染的鼠标，或可达30-40×10 6中性粒细胞/感染小鼠）纯（80 -95％）嗜中性粒细胞&gt; 95％的存活率，从骨髓。此方法使用市售HISTOPAQUE，密度梯度细胞分离配给介质组成的聚蔗糖和泛影葡胺钠，分离从小鼠骨髓中的嗜中性粒细胞。此方法产生的显着更大的每只小鼠的嗜中性粒细胞的数目相比，血液或腹膜腔，它可以被用来收集在稳定状态或感染后的小鼠嗜中性粒细胞，它是比较容易的层相比，使用不连续的密度梯度离心法Percoll密度梯度组成的55％/ 65％/ 75％的Percoll 9在PBS中。此外，收集纯净的中性粒细胞所需的时间和资源相比显着下降，中性粒细胞隔离，利用荧光激活细胞分选。此外，因为这种方法不涉及免疫磁性富集步骤，更具成本效益的，它避免了细胞暴露磁性柱和抗体，从而减少中性粒细胞活化的可能性。 在除了执行功能隔离neutroph研究中ILS 体外和细胞过继转移到受体小鼠，该协议还描述了一种方法，用于标识分离的嗜中性粒细胞，使用不同的CellTracker染料。从不同遗传背景的小鼠中性粒细胞的鉴别标签跟踪转移的嗜中性粒细胞在受体小鼠的组织，采用流式细胞仪，它可以提供机械洞察力贩运中性粒细胞从血液中特定基因的直接作用，可适应在竞争激烈的复育研究进入靶器官发炎6。

<table border="1">
 <tbody>
 <tr>
 <td>Name of Reagent/Material</td>
 <td>Company</td>
 <td>Catalog Number</td>
 <td>Comments</td>
 </tr>
 <tr>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 <td>Reagents</td>
 </tr>
 <tr>
 <td>RPMI 1640 1X with L-glutamine and 25 mM HEPES</td>
 <td>Cellgro</td>
 <td>10-041-CV</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Fetal Bovine Serum (Heat inactivated)</td>
 <td>GemCell</td>
 <td>100500</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Penicillin/Streptomycin (10,000 units penicillin / 10,000 mg/ml strep)</td>
 <td>GIBCO</td>
 <td>15140</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>0.5 M EDTA</td>
 <td>Quality Biological Inc</td>
 <td>351-027-101</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>0.2% and 1.6% sodium chloride</td>
 <td>JT Baker</td>
 <td>3624</td>
 <td>Sodium chloride solutions prepared using distilled water and sterile filtered</td>
 </tr>
 <tr>
 <td>Sterile filtered Histopaque 1077</td>
 <td>Sigma</td>
 <td>10771</td>
 <td>Histopaque 1077 needs to be brought to 18-26 &deg;C before use</td>
 </tr>
 <tr>
 <td>Sterile filtered Histopaque 1119</td>
 <td>Sigma</td>
 <td>11191</td>
 <td>Histopaque 1119 needs to be brought to 18-26 &deg;C before use</td>
 </tr>
 <tr>
 <td>Phosphate Buffered Saline (PBS) without Calcium and Magnesium</td>
 <td>Cellgro</td>
 <td>210-40-CV</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Ethyl Alcohol (200 proof)</td>
 <td>The Warner Graham Company</td>
 <td>64-17-5</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>CellTracker Green (CMFDA, 5-Chloromethylfluorescein Diacetate)</td>
 <td>Invitrogen</td>
 <td>C-7025</td>
 <td>Make stock solutions of 10 mM in DMSO, aliquot and store at -80 &deg;C</td>
 </tr>
 <tr>
 <td>CellTracker Orange (CMTMR, 5-(and 6)-4-Chloromethyl Benzoyl Amino Tetramethylrhodamine)</td>
 <td>Invitrogen</td>
 <td>C-2927</td>
 <td>Make stock solutions of 10 mM in DMSO, aliquot and store at -80 &deg;C</td>
 </tr>
 <tr>
 <td>Anti-mouse CD45 (Clone 30-F11)</td>
 <td>eBioscience</td>
 <td>170451-82</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Anti-mouse Ly6G (Clone 1A8)</td>
 <td>BD Pharmingen</td>
 <td>551461</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Anti-mouse Ly6G (Clone 1A8)</td>
 <td>BD Pharmingen</td>
 <td>560599</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Anti-mouse CD11b (Clone M1/70)</td>
 <td>eBioscience</td>
 <td>47-0112-82</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Anti-mouse CD16/CD32 (Mouse BD Fc Block)</td>
 <td>BD Pharmingen</td>
 <td>553141</td>
 <td>Use at 1:100 dilution</td>
 </tr>
 <tr>
 <td>LIVE/DEAD Fixable Blue Dead Cell Stain Kit</td>
 <td>Molecular Probes</td>
 <td>L-23105</td>
 <td>Use at 1:1000 dilution</td>
 </tr>
 <tr>
 <td>Dimethyl Sulfoxide (DMSO)</td>
 <td>Sigma</td>
 <td>67-68-5</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Paraformaldehyde Solution, 4% in PBS</td>
 <td>USB Corporation</td>
 <td>19943</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 <td>Table 1. Reagents for isolation, labeling and tracking of adoptively transferred neutrophils.</td>
 </tr>
 <tr>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 <td>Materials</td>
 </tr>
 <tr>
 <td>C57BL/6 mice</td>
 <td>Taconic</td>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>15 ml centrifuge tubes</td>
 <td>Corning</td>
 <td>430053</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>50 ml centrifuge tubes</td>
 <td>BD Falcon</td>
 <td>352070</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>25 ml serological pipettes</td>
 <td>Celltreat</td>
 <td>229225B</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>10 ml serological pipettes</td>
 <td>Celltreat</td>
 <td>229210B</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>5 ml serological pipettes</td>
 <td>Celltreat</td>
 <td>229205B</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Pasteur pipettes (3 ml)</td>
 <td>BD Falcon</td>
 <td>357575</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>12 ml syringes</td>
 <td>Kendall monoject</td>
 <td>512878</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>25 G x 5/8 in. Needles (precision glide needles)</td>
 <td>BD</td>
 <td>305122</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>100 mm cell strainers</td>
 <td>BD Falcon</td>
 <td>352360</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Bactericidal Petri dishes</td>
 <td>BD Falcon</td>
 <td>351029</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Combitips Plus Biopur</td>
 <td>Eppendorf</td>
 <td>2249608-5</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Mouse dissecting instruments (Scissors, forceps, scalpel)</td>
 <td>Biomedical Research Instruments</td>
 <td>10-2300, 10-2165, 25-1200, 26-1000</td>
 <td>Instruments sterilized prior to use</td>
 </tr>
 <tr>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 <td>Equipment</td>
 </tr>
 <tr>
 <td>Tissue culture hood</td>
 <td>The Baker Company</td>
 <td>SG403</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>Refrigerated centrifuge</td>
 <td>Thermo Fischer Scientific</td>
 <td>75004521</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>37 &deg;C shaking water bath</td>
 <td>Thermo Fischer Scientific</td>
 <td>3166721</td>
 <td>&nbsp;</td>
 </tr>
 <tr>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 <td>&nbsp;</td>
 <td>Table 2. List of Materials and Equipment used in this protocol.</td>
 </tr>
 </tbody>
</table>

isolation, purification and labeling of mouse bone marrow neutrophils for functional studies and adoptive transfer experiments

嗜中性粒细胞是关键的先天免疫系统的效应细胞。他们正在迅速招募急性炎症部位，发挥保护或致病作用取决于炎症环境。然而，尽管中性粒细胞免疫力，详细了解调解中性粒细胞的效应，在不同的传染性疾病和炎症性疾病的免疫病理作用的分子因素，不可或缺的作用仍然缺乏，部分原因是由于其半衰期短，处理这些困难细胞和缺乏可靠的实验协议，获得足够数量的下游功能研究和中性粒细胞过继转移实验。因此，操作简单，快速，经济，可靠的方法是非常可取的收获足够数量的鼠标中性粒细胞的吞噬功能，杀灭，细胞因子的产生，脱粒和贩运等功能评估。为此目的，我们提出了可再现的密度梯度离心法为基础的协议，它可以适应于任何的实验室分离大量的中性粒细胞从小鼠骨髓中的具有高纯度和活力。此外，我们提出了一个简单的协议，使用CellTracker染料标记隔离中性粒细胞，然后可以过继转移到受体小鼠在一些组织和跟踪至少4个小时的转移后，使用流式细胞仪。使用这种方法，可以成功地采用了差异化标记的嗜中性粒细胞从野生型和基因缺陷小鼠不同CellTracker染料，执行竞争力的评价贩卖中性粒细胞从血液中特定基因的直接作用到在体内靶组织的再增殖研究。

这个协议被优化用于从小鼠骨髓细胞的收获和随后的分离的嗜中性粒细胞通过密度梯度离心使用市售HISTOPAQUE细胞分离介质从这些细胞。嗜中性粒细胞分离，使用这种方法可用于多种下游功能性研究体外受体小鼠过继转移实验中。 未受感染8-12周龄C57BL / 6小鼠双侧股骨和胫骨的骨髓细胞，每收集典型的产量为60-80万〜〜94-98％具有可行性细胞。下行密度梯度离心这些细胞?...

Watch this Scientific Journal Video about Isolation, Purification and Labeling of Mouse Bone Marrow Neutrophils for Functional Studies and Adoptive Transfer Experiments at JoVE.com

Isolation, Purification and Labeling of Mouse Bone Marrow Neutrophils for Functional Studies and Adoptive Transfer Experiments

我们描述了一个协议，用于分离和纯化从小鼠骨髓中的嗜中性粒细胞通过密度梯度离心和嗜中性粒细胞标记使用CellTracker染料。这是一个简单，快速，重复性好，经济的方法获得大量的嗜中性粒细胞的下游功能研究或过继转移和跟踪实验。

功能研究和过继转移实验小鼠骨髓嗜中性粒细胞的分离，纯化及贴标

Neutrophils are critical effector cells of the innate immune system. They are rapidly recruited at sites of acute inflammation and exert protective or ...

Research

JoVE Journal

Immunology and Infection

Experimental Metastasis and CTL Adoptive Transfer Immunotherapy Mouse Model

实验转移和CTL过继转移免疫小鼠模型

Experimental metastasis mouse model is a simple and yet physiologically relevant metastasis model. The tumor cells are injected intravenously (i.v) into ...

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免疫与感染

Generation and Labeling of Murine Bone Marrow-derived Dendritic Cells with Qdot Nanocrystals for Tracking Studies

小鼠骨髓来源的树突状细胞与跟踪研究的Qdot纳米晶的生成及标签

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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 ...

Isolation and Intravenous Injection of Murine Bone Marrow Derived Monocytes

隔离和小鼠骨髓的静脉注射单核细胞衍生

As a subtype of leukocytes and progenitors of macrophages, monocytes are involved in many important processes of organisms and are often the subject of ...

Tracking Mouse Bone Marrow Monocytes In Vivo

Tracking Mouse Bone Marrow Monocytes In Vivo

跟踪小鼠骨髓单核细胞<em&gt;在体内</em

Real time multiphoton imaging provides a great opportunity to study cell trafficking and cell-to-cell interactions in their physiological 3-dimensionnal ...

Murine Hind Limb Long Bone Dissection and Bone Marrow Isolation

鼠后肢长骨解剖和骨髓隔离

Investigation of the bone and the bone marrow is critical in many research fields including basic bone biology, immunology, hematology, cancer metastasis, ...

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 ...

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 ...