Name: in vivo 19f mri for cell tracking
Uploaded: 2013-11-25T21:10:00
Description: Watch this Scientific Journal Video about In vivo 19F MRI for Cell Tracking at JoVE.com

我们要感谢纳丁·海恩和阿伦德Heerschap的宝贵援助。这项工作得到了再生医学的荷兰研究所（NIRM，FES0908），欧盟第七框架计划ENCITE（医学-F5-2008-201842）和TargetBraIn（医学-F2-2012-279017），从大众汽车基金会的资助（支持的I/83 443），荷兰科学研究组织（VENI 700.10.409和VIDI 917.76.363），ERC（高级格兰特269019）和Radboud大学Nijmegen医学中心（AGIKO-2008-2-4）。

注：所有的实验和程序涉及动物，必须进行按照相关的道德准则和符合标准的动物护理和人性化的要求。 1。细胞标记（与共同孵育标准协议） <ol><li>添加19女性标签8未成熟DC在4 mg/10 6细胞浓度（2毫升培养基）。轻轻摇晃混合。 </li><li>孵育的细胞与标签的成熟和收获的DC前3天。 </li><li>收集区议会，并在PBS冲洗至少3倍去除多余的标签。计数细胞。 </l...

<ol>
	<li>Srinivas, M., 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=Imaging+of+cellular+therapies.">Imaging of cellular therapies.</a> Adv. Drug Deliv. Rev. 62, 1080-1093 (2010).</li><li>de Vries, I. J., 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=Magnetic+resonance+tracking+of+dendritic+cells+in+melanoma+patients+for+monitoring+of+cellular+therapy.">Magnetic resonance tracking of dendritic cells in melanoma patients for monitoring of cellular therapy.</a> Nat. Biotechnol. 23, 1407-1413 (2005).</li><li>Srinivas, M., Heerschap, A., Ahrens, E. T., Figdor, C. G., de Vries, I. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=MRI+for+quantitative+in+vivo+cell+tracking.">MRI for quantitative in vivo cell tracking.</a> Trends Biotechnol. 28, 363-370 (2010).</li><li>Ruiz-Cabello, J., Barnett, B. P., Bottomley, P. A., Bulte, J. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorine+(19F)+MRS+and+MRI+in+biomedicine.">Fluorine (19F) MRS and MRI in biomedicine.</a> NMR Biomed. 24, 114-129 (2011).</li><li>Stoll, G., Basse-Lusebrink, T., Weise, G., Jakob, P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visualization+of+inflammation+using+19F-magnetic+resonance+imaging+and+perfluorocarbons.">Visualization of inflammation using 19F-magnetic resonance imaging and perfluorocarbons.</a> Wires Nanomed. Nanobiol. 4, 438-447 (2012).</li><li>Srinivas, M., Boehm-Sturm, P., Figdor, C. G., de Vries, I. J., Hoehn, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Labeling+cells+for+in+vivo+tracking+using+(19)F.">Labeling cells for in vivo tracking using (19)F.</a> MRI. Biomaterials. 33, 8830-8840 (2012).</li><li>Ahrens, E. T., Flores, R., Xu, H., Morel, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+imaging+platform+for+tracking+immunotherapeutic+cells.">In vivo imaging platform for tracking immunotherapeutic cells.</a> Nat. Biotechnol. 23, 983-987 (2005).</li><li>Srinivas, M., 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=Customizable,+multi-functional+fluorocarbon+nanoparticles+for+quantitative+in+vivo+imaging+using+19F+MRI+and+optical+imaging.">Customizable, multi-functional fluorocarbon nanoparticles for quantitative in vivo imaging using 19F MRI and optical imaging.</a> Biomaterials. 31, 7070-7077 (2010).</li><li>Boehm-Sturm, P., Mengler, L., Wecker, S., Hoehn, M., Kallur, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vivo+tracking+of+human+neural+stem+cells+with+19F+magnetic+resonance+imaging.">In vivo tracking of human neural stem cells with 19F magnetic resonance imaging.</a> PLoS One. 6, e29040 (2011).</li><li>Srinivas, M., 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=In+vivo+cytometry+of+antigen-specific+t+cells+using+(19)F.">In vivo cytometry of antigen-specific t cells using (19)F.</a> MRI. Magn. Reson. Med. , (2009).</li><li>Srinivas, M., Morel, P. A., Ernst, L. A., Laidlaw, D. H., Ahrens, E. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Fluorine-19+MRI+for+visualization+and+quantification+of+cell+migration+in+a+diabetes+model.">Fluorine-19 MRI for visualization and quantification of cell migration in a diabetes model.</a> Magn. Reson. Med. 58, 725-734 (2007).</li><li>Mangala Srinivas, E. T. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cellular+labeling+and+quantification+for+nuclear+magnetic+resonance+techniques.">Cellular labeling and quantification for nuclear magnetic resonance techniques.</a> US patent. , (2007).</li><li>Boehm-Sturm, P., Pracht, E. D., Aswendt, M., Henn, N., Hoehn, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Proceedings of the International Society for Magnetic Resonance in Medicine. , (2012).</li><li>Insko, E. K., Bolinger, L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mapping+of+the+Radiofrequency+Field.">Mapping of the Radiofrequency Field.</a> J. Magn. Res. A. 103, 82-85 (1993).</li><li>Haacke, E. M., Brown, R. W., Thompson, M. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=.">.</a> Magnetic resonance imaging: physical principles and sequence design. , (1999).</li><li>Scheffler, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+pictorial+description+of+steady-states+in+rapid+magnetic+resonance+imaging.">A pictorial description of steady-states in rapid magnetic resonance imaging.</a> Concepts Magn. Res. 11, 291-304 (1999).</li><li>Firbank, M. J., Coulthard, A., Harrison, R. M., Williams, E. 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+comparison+of+two+methods+for+measuring+the+signal+to+noise+ratio+on+MR+images.">A comparison of two methods for measuring the signal to noise ratio on MR images.</a> Phys. Med. Biol. 44, 261-264 (1999).</li><li>de Chickera, S. N., 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=Labelling+dendritic+cells+with+SPIO+has+implications+for+their+subsequent+in+vivo+migration+as+assessed+with+cellular+MRI.">Labelling dendritic cells with SPIO has implications for their subsequent in vivo migration as assessed with cellular MRI.</a> Contrast Media Mol. Imaging. 6, 314-327 (2011).</li></ol>

这里所概述的协议描述了用于在体内 19女性 MRI小区追踪的一般方法。尽管所描述的共培养方法，有用于与19女性剂标记细胞数不同的协议。然而，共培养是最常用的，并且可以通过加入转染试剂6的进一步优化，例如 。实际的细胞标记协议将取决于细胞类型。唯一密钥标记的步骤在这里描述的协议。通常，标签的制备和加入到细胞中的选定时间范围从几个小?...

体内细胞跟踪是用于细胞疗法1的优化和监测至关重要。由于其非侵入性的性质，成像提供了极好的机会来监测细胞在体内 。磁共振成像（MRI）是独立的电离辐射，并且允许一个优越的成像分辨率和固有软组织的对比。基于MRI的小区追踪已经用于临床跟踪的树突状细胞中的黑色素瘤患者2。传统的临床MRI进行的1小时核，目前在用流动清水组织中。另外，也可以进行MRI上的其他活性核，如13 C，19 F和23的Na。然而，只有19女性 MRI已成功应用于体内细胞示踪因为它提供了1后H的最高灵敏度在组织中缺乏的MRI检测的内源性19女性允许高信号选择性的检测外源性19 F对比剂，并允许直接从图像数据的氟浓度进行定量。有关19女性 MRI的详细讨论，请参见3-5。与19女性 MRI检查的一个关键问题是需要开发和优化适合19女性电池标签，虽然有几个标签已被开发，以实现多式联运代理6的趋势。 我们在这里描述的协议是由我们的团体7-9，其中所描述的体内定量19女性基于MRI的细胞跟踪10,11的第一篇文章的基础上的研究。使用19女性 MRI小区追踪的一般方法概述于图1中 。我们描述了一个通用协议的树突状细胞（DCs）使用一个特制的氟碳造影剂8的标记和成像。成像协议是普遍适用于不同细胞类型，标签和动物模型。该这里描述的细胞类型和动物模型应该只作为例子，并且因此我们不提供对细胞的分离和标识细节，而是着眼于成像协议。修改将是必要为每个标签，细胞类型，动物模型和成像设置，并且这些可以在文献中找到，或可能需要由研究者进行优化。一些常见的修改都包括在讨论。

<table border="1">
		<tbody>
		 <tr>
			<td></td>
			<td></td>
			<td></td>
			<td>REAGENTS</td>
		 </tr>
		 <tr>
			<td>PBS</td>
			<td>Sigma-Aldrich</td>
			<td>MFCD00131855</td>
			<td></td>
		 </tr>
		 <tr>
			<td>TFA</td>
			<td>Sigma-Aldrich</td>
			<td>76-05-1 </td>
			<td></td>
		 </tr>
		 <tr>
			<td>Ketamine (Ketavet)</td>
			<td>Pfizer</td>
			<td>778-551</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Xylazine (Rompun)</td>
			<td>Bayer</td>
			<td>QN05 cm92</td>
			<td></td>
		 </tr>
		 <tr>
			<td>Ophtosan </td>
			<td>Produlab Pharma</td>
			<td>2702</td>
			<td>eye ointment</td>
		 </tr>
		 <tr>
			<td></td>
			<td></td>
			<td></td>
			<td>Material name </td>
		 </tr>
		 <tr>
			<td>MRI scanner</td>
			<td>Bruker Biospec</td>
			<td></td>
			<td></td>
		 </tr>
		 <tr>
			<td>NMR spectrometer</td>
			<td>Bruker Biospec</td>
			<td></td>
			<td></td>
		 </tr>
		</tbody>
 </table>

in vivo 19f mri for cell tracking

在体内 19女性 MRI检查可定量细胞追踪，而无需使用电离辐射。它是一个可以应用到人类的非侵入性技术。在这里，我们描述了一种通用协议，用于细胞标记，成像和图像处理。该技术适用于各种类型的细胞和动物模型中，虽然在这里，我们专注于一个典型的小鼠模型来跟踪小鼠的免疫细胞。细胞标记的最重要的问题进行了描述，因为这些是适用于所有型号。同样的，关键的成像参数都列出来，虽然细节将根据MRI系统和个人设置的不同而不同。最后，我们有定量的图像处理协议。变化对于这一点，并且协议的其他部分，被评估的讨论部分。根据此处所描述的具体步骤，用户将需要适应的协议为每个特定的细胞类型，细胞标记，动物模型和成像设置。注意，在protocol还可以适于人用的，只要临床限制得到满足。

这里，我们表明，涉及19 F-标记的细胞归巢至引流淋巴结的转移协议的典型结果。 图2示出了使用TFA参考10 6标记细胞的19 F NMR光谱。成像设置进行在协议（ 图3）中所述。对于体内成像，我们使用了一个参考由同一标签的用于细胞放置在鼠标的脚之间的密封圆柱体。一位代表处理后的图像示于图4。通过应用在7.2节中描述的协议，...

Watch this Scientific Journal Video about In vivo 19F MRI for Cell Tracking at JoVE.com

In vivo 19F MRI for Cell Tracking

我们描述了一种通用协议，用于在小鼠模型中具有活体外标记的细胞用MRI 体内细胞的跟踪。包含一个典型的协议，用于细胞标记，图像采集处理和量化。

在体内 19女性 MRI对细胞跟踪

In vivo 19F MRI allows quantitative cell tracking without the use of ionizing radiation. It is a noninvasive technique that can be applied to humans. ...

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