Name: multimodal imaging of stem cell implantation in the central nervous system of mice
Uploaded: 2012-06-13T13:00:00
Description: Watch this Scientific Journal Video about Multimodal Imaging of Stem Cell Implantation in the Central Nervous System of Mice at JoVE.com

<p class="jove_content"&gt;作者的工作得到了转炉研究资助ID-2006年的安特卫普大学（PPO和AVDL授予），研究资助G.0136.11和G.0130.11（授予AVDL，ZB和PPO）和1.5.021.09。 N.00（授予PPO）的科学研究，法兰德斯（FWO Vlaanderen，比利时），服务营办商研究资助内陆水运-60838：BRAINSTIM佛兰德科技研究所（ZB和AVDL授予），在基金1玛土撒拉的研究经费从弗拉芒政府（授予ZB型），部分由EC-FP6的NOE迪米（LSHB-CT-2005-512146），EC-FP6的NOE EMIL（LSHC-CT-2004-503569的部分）间大学景点波兰人IUAP-NIMI-P6/38（授予AVDL）。纳塔莉德Vocht FWO-Vlaanderen持有博士学位的奖学金。彼得Ponsaerts是的FWO-Vlaanderen的博士后研究员。</p

1。细胞的制备<ol><li>应开始实验使用体外培养干细胞基因工程表达的荧光素酶和绿色荧光蛋白记者蛋白质的人群。在这里，我们使用荧光素酶/ EGFP表达小鼠骨髓来源的基质细胞（BMSC-Luc/eGFP）以前Bergwerf 等。2，5。 </li><li>细胞标记，在8×10 5％的T75培养瓶中的细胞密度在15毫升完整的扩展介质（CEM），辅以1微克/毫升Puromycine板BMSC-Luc/eGFP细胞前两天。 </li><li>让细胞成长为48小?...

<ol>
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在这份报告中，我们描述了一个在免疫小鼠中枢神经系统的细胞植入的详细描述。（BLI，MRI和组织学）三个互补的成像方式的组合优化的协议。记者细胞的基因标记，记者萤火虫荧光素酶和绿色荧光蛋白基因，并直接与GB的MPIO细胞标记基因改造的基础上，组合会导致体内的干细胞移植在准确的评估。 粒子标记的骨髓间充质干细胞，GB的的MPIO粒径组合与阴离子表?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td> IMDM培养</td><td>龙沙</td><td> BE12-722F </td><td>细胞的生长介质的CEM组件</td></tr><tr><td>胎牛血清</td><td> Gibco公司</td><td> 10270-106 </td><td>细胞的生长介质的CEM组件</td></tr><tr><td>马血清</td><td> Gibco公司</td><td> 1605-122 </td><td>细胞的生长介质的CEM组件</td></tr><tr><td>青霉素链霉素</td><td> Gibco公司</td><td> 15140 </td><td>细胞的生长介质的CEM组件</td></tr><tr><td> fungizone </td><td> Gibco公司</td><td> 15290-018 </td><td>细胞的生长介质的CEM组件</td></tr><tr><td> PBS </td><td> Gibco公司</td><td> 14190 </td><td></td></tr><tr><td> puromycine </td><td> invivogen </td><td> ANT-PR-1 </td><td></td></tr><tr><td>胰蛋白酶</tD&gt; <td> Gibco公司</td><td> 25300 </td><td></td></tr><tr><td> GB的MPIO的</td><td>刘海实验室</td><td> ME04F/7833 </td><td></td></tr><tr><td>的D-荧光素</td><td> Promega公司</td><td> E1601 </td><td></td></tr><tr><td>氯胺酮（Ketalar） </td><td>辉瑞公司</td><td></td><td></td></tr><tr><td>甲苯噻嗪（Rompun） </td><td>拜耳医疗保健</td><td></td><td></td></tr><tr><td>异氟醚</td><td> isoflo </td><td> 05260-05 </td><td></td></tr><tr><td> 0.9％NaCl溶液</td><td>巴克斯特</td><td></td><td></td></tr><tr><td>多聚甲醛</td><td>默克公司</td><td> 1.04005.1000 </td><td></td></tr><tr><td>蔗糖</td><td> applichem </td><td> A1125 </td><td></td></tr><tr><td>微量注射泵</td><td>川崎病科学</td><td> KDS100 </td><td></td></tr><tr><td>光子成像</td><td> biospace实验室</td><td></td><td></td></tr><tr&gt; <td> 9.4T议员扫描器</td><td>布鲁克BioSpin </td><td> biospec 94/20的USR </td><td></td></tr><tr><td> BX51显微镜</td><td>奥林巴斯</td><td> BX51 </td><td></td></tr><tr><td> mycrom航模低温恒温器</td><td> prosan </td><td> HM525 </td><td></td></tr><tr><td>注射器</td><td>汉密尔顿</td><td> 7635-01 </td><td></td></tr><tr><td> 30号针头</td><td>汉密尔顿</td><td> 7762-03 </td><td></td></tr><tr><td>照片Vision软件</td><td> biospace实验室</td><td></td><td></td></tr><tr><td> m3vision软件</td><td> biospace实验室</td><td></td><td></td></tr><tr><td> paravision 5.1软件</td><td>布鲁克BioSpin </td><td></td><td></td></tr><tr><td>阿米拉4.0软件</td><td> Visage的影像</td><td></td><td></td></tr></tbody></table>

multimodal imaging of stem cell implantation in the central nervous system of mice

在过去十年中，造血干细胞移植已越来越为原发性或继发多种疾病，在临床前和临床研究的治疗方法的兴趣。然而，迄今为止，有关功能的结果和/或组织再生以下干细胞移植的结果是多种多样的。一般来说，临床受益观察没有深刻理解底层机制（S）1。因此，多的努力已导致不同的分子成像的发展，以监察其最终目的的干细胞移植，以准确评估移植的干细胞和/或他们的微环境的生存，命运和生理。分子成像确定一个或多个参数的变化可能与观察到的临床效果。在此背景下，我们的研究侧重于生物发光成像的结合使用。（BLI），磁共振成像（MRI）和病理analysis到评价干细胞移植。 ，劳保局常用非侵入性细胞跟踪和监视细胞的生存时间，移植后2-7细胞表达荧光素酶报告基因的生化反应的基础上能够发出光以下的相互作用，其基板（例如D-荧光素），8，9。另一方面，MRI是一种非侵入性的技术，这是临床上适用10，可用于精确定位，具有很高的分辨率11-15细胞移植，但其灵敏度高，取决于细胞标记后生成的磁共振造影剂的对比。最后，验尸的病理分析的首选方法，以验证获得最高的分辨率和灵敏度的非侵入性技术的研究成果。此外终点的病理分析，让我们进行详细的表型分析移植的细胞和/或周围组织，BASED上使用荧光记者蛋白质和/或直接与特异性抗体细胞标记。 总之，我们在这里直观地展现劳保局，MRI和组织学的互补性，解开不同的干细胞和/或环境相关的特征后，干细胞移植在小鼠中枢神经系统。作为一个例子，骨髓基质细胞，基因工程表达增强型绿色荧光蛋白（EGFP）和萤火虫荧光素酶（fLuc），和蓝色荧光微米大小的氧化铁粒子（MPIOs）标记，将嫁接中枢神经系统免疫能力的小鼠和成果将监测劳保局，MRI和组织学（ 图1）。

Watch this Scientific Journal Video about Multimodal Imaging of Stem Cell Implantation in the Central Nervous System of Mice at JoVE.com

Multimodal Imaging of Stem Cell Implantation in the Central Nervous System of Mice

本文介绍了优化多式联运成像啮齿动物脑细胞的使用移植的事件序列：（i）在活体动物和磁共振成像，及（ii）验尸的病理分析。结合在一个单一的动物允许这些成像分辨率高，灵敏度和特异性细胞移植评估。

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标记的单细胞在中枢神经系统<em&gt;果蝇</em

In this  article we describe how to individually label neurons in the embryonic CNS of Drosophila melanogaster by juxtacellular injection of the ...

Direct Intraventricular Delivery of Drugs to the Rodent Central Nervous System

啮齿动物的中枢神经系统药物直接脑室交货

Due to an inability to cross the blood brain barrier, certain drugs need to be directly delivered into the central nervous system (CNS). Our lab focuses ...

In vivo Interrogation of Central Nervous System Translatome by Polyribosome Fractionation

In vivo Interrogation of Central Nervous System Translatome by Polyribosome Fractionation

在中枢神经系统Translatome由多核糖体分馏体内拷问

Multiple processes are involved in gene expression including transcription, translation and stability of mRNAs and proteins. Each of these steps are ...

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

In vivo Optogenetic Stimulation of the Rodent Central Nervous System

在啮齿动物中枢神经系统的体内光遗传学刺激

The ability to probe defined neural circuits in awake, freely-moving animals with cell-type specificity, spatial precision, and high temporal resolution ...

Functional Evaluation of Biological Neurotoxins in Networked Cultures of Stem Cell-derived Central Nervous System Neurons

生物神经毒素在茎的网络文化功能评估细胞衍生的中枢神经系统神经元

Therapeutic and mechanistic studies of the presynaptically targeted clostridial neurotoxins (CNTs) have been limited by the need for a scalable, ...

In Situ Ca2+ Imaging of the Enteric Nervous System

In Situ Ca2+ Imaging of the Enteric Nervous System

在肠神经系统的原位 钙成像

Reflex behaviors of the intestine are controlled by the enteric nervous system (ENS). The ENS is an integrative network of neurons and glia in two ...

Optical Clearing of the Mouse Central Nervous System Using Passive CLARITY

鼠标中枢神经系统的光透明使用被动CLARITY

Traditionally, tissue visualization has required that the tissue of interest be serially sectioned and imaged, subjecting each tissue section to unique ...

Electrophysiological Recording of The Central Nervous System Activity of Third-Instar Drosophila Melanogaster

Electrophysiological Recording of The Central Nervous System Activity of Third-Instar  Drosophila Melanogaster

第三星果蝇中枢神经系统活动的电生理记录

The majority of the currently available insecticides target the nervous system and genetic mutations of invertebrate neural proteins oftentimes yield ...