Name: electric field-controlled directed migration of neural progenitor cells in 2d and 3d environments
Uploaded: 2012-02-16T12:00:00
Description: Watch this Scientific Journal Video about Electric Field-controlled Directed Migration of Neural Progenitor Cells in 2D and 3D Environments at JoVE.com

这项工作得到了英国皇家学会的统一报告格式授予UF051616，英国和欧洲研究理事会英镑赠款243261到BS。也支持加州理工学院的再生医学批RB1-01417在锰锌实验室工作。

1。神经祖细胞的分离<ol><li> E14-16小鼠在寒冷的DMEM/F12培养基进行剖析，从全脑。删除所有脑膜转移到35毫米的培养皿下解剖显微镜和大脑。 </li><li>用细镊子机械分解成组织碎片的大脑和他们转移到15 ml管，然后离心3分钟的样本，以消除碎片在800转。 </li><li>加入DMEM/F12培养液含有碱性成纤维细胞生长因子和表皮生长因子和磨碎用1毫升吸管。 </li><li>通过一个细胞过滤细胞悬液，以获得单细胞悬液?...

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	<li>Huttenlocher, A., Horwitz, A. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Wound+healing+with+electric+potential.">Wound healing with electric potential.</a> N. Engl. J. Med. 356, 303-303 (2007).</li><li>McCaig, C. D., Rajnicek, A. M., Song, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Controlling+cell+behavior+electrically:+current+views+and+future+potential.">Controlling cell behavior electrically: current views and future potential.</a> Physiol. Rev. 85, 943-943 (2005).</li><li>Borgens, R. B., Jaffe, L. F., Cohen, M. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Large+and+persistent+electrical+currents+enter+the+transected+lamprey+spinal+cord.">Large and persistent electrical currents enter the transected lamprey spinal cord.</a> PNAS. 77, 1209-1209 (1980).</li><li>Shapiro, S., Borgens, R., Pascuzzi, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Oscillating+field+stimulation+for+complete+spinal+cord+injury+in+humans:+a+phase+1+trial.">Oscillating field stimulation for complete spinal cord injury in humans: a phase 1 trial.</a> J. Neurosurg. Spine. 2, 3-3 (2005).</li><li>Zhao, M., Song, B., Pu, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrical+signals+control+wound+healing+through+phosphatidylinositol-3-OH+kinase-gamma+and+PTEN.">Electrical signals control wound healing through phosphatidylinositol-3-OH kinase-gamma and PTEN.</a> Nature. 442, 457-457 (2006).</li><li>Yao, L., Shanley, L., McCaig, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small+applied+electric+fields+guide+migration+of+hippocampal+neurons.">Small applied electric fields guide migration of hippocampal neurons.</a> J. Cell. Physiol. 216, 527-527 (2008).</li><li>Li, L., El-Hayek, Y. H., Liu, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Direct-current+electrical+field+guides+neuronal+stem&#47;progenitor+cell+migration.">Direct-current electrical field guides neuronal stem&#47;progenitor cell migration.</a> Stem Cells. 26, 2193-2193 (2008).</li><li>Meng, X., Arocena, M., Penninger, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=PI3K+mediated+electrotaxis+of+embryonic+and+adult+neural+progenitor+cells+in+the+presence+of+growth+factors.">PI3K mediated electrotaxis of embryonic and adult neural progenitor cells in the presence of growth factors.</a> Exp. Neurol. 227, 210-210 (2011).</li><li>Bonnici, B., Kapfhammer, J. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bone+marrow+stromal+cells+promote+neurite+extension+in+organotypic+spinal+cord+slice:+significance+for+cell+transplantation+therapy.">Bone marrow stromal cells promote neurite extension in organotypic spinal cord slice: significance for cell transplantation therapy.</a> Neurorehabil. Neural Repair. 22, 447-447 (2008).</li><li>Shichinohe, H., Kuroda, S., Tsuji, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bone+marrow+stromal+cells+promote+neurite+extension+in+organotypic+spinal+cord+slice:+significance+for+cell+transplantation+therapy.">Bone marrow stromal cells promote neurite extension in organotypic spinal cord slice: significance for cell transplantation therapy.</a> Neurorehabil. Neural Repair. 22, 447-447 (2008).</li><li>Song, B., Gu, Y., Pu, j. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Application+of+direct+current+electric+fields+to+cells+and+tissues+in+vitro+and+modulation+of+wound+electric+field+in+vivo.">Application of direct current electric fields to cells and tissues in vitro and modulation of wound electric field in vivo.</a> Nature Protocol. 2, 1479-1479 (2007).</li></ol>

5,11以前的研究基础上，我们所使用的协议。使用这些方法，稳定的文化和电目前的条件，可同时申请通过琼脂桥梁，Steinberg的解决方案，Ag / AgCl电极，标准化和精确的尺寸定制设计electrotactic的商会，培养细胞或片的EF维护。钱伯斯的深度可以调整，以适应不同的样品厚度11，在细胞的情况下，室的大小可以修改，以适应但是许多细胞（如Western blot分析需要很多的细胞）。我们已?...

<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>基本的FGF-重组人</td><td> Invitrogen公司</td><td> PHG0026 </td><td> 20毫微克/毫升</td></tr><tr><td>表皮生长因子重组人</td><td> Invitrogen公司</td><td> PHG0311 </td><td> 20毫微克/毫升</td></tr><tr><td> N2 - 补充液体（100X） </td><td> Invitrogen公司</td><td> 02048 </td><td></td></tr><tr><td> DMEM/F12培养基（高糖） </td><td> Invitrogen公司</td><td> 31330-095 </td><td></td></tr><tr><td>聚-D-赖氨酸</td><td> Millipore公司</td><td> -003-E的</td><td></td></tr><tr><td>自然科学小鼠层粘连蛋白</td><td> Invitrogen公司</td><td> 23017-015 </td><td></td></tr><tr><td>生长因子减少基底膜基质（Matr​​igel的） </td><td> BD公司</TD&gt; <td> 354230 </td><td></td></tr><tr><td> HEPES缓冲</td><td> Gibco公司</td><td> 15630 </td><td></td></tr><tr><td> McIlwain组织菜刀</td><td>小数怕长计实验室工程有限公司</td><td> TC752-PD </td><td></td></tr><tr><td>道康宁高真空硅脂</td><td> Sigma-Aldrich公司</td><td> Z273554 </td><td></td></tr></tbody></table>

electric field-controlled directed migration of neural progenitor cells in 2d and 3d environments

内源性电场（EFS） 在体内自然发生和组织/器官的发育和再生过程中发挥关键作用，包括中枢神经系统1,2。这些内源性的EF所产生的细胞与细胞和组织的电阻相结合的离子运输调控。据报道，应用EF治疗可以促进脊髓损伤，动物和人类的3,4功能修复。 EF导向细胞迁移，特别是已被证实在多种类型的细胞5,6，包括神经祖细胞（筹备）7,8。直流电（DC）EF的应用是不是在大多数实验室常用的技术。我们已经介绍了详细的协议，为直流EF的应用细胞和组织培养，以前5,11。在这里，我们提出了一个标准方法的视频示范的基础上计算磁场强度成立2D 1ð3D环境，为筹备和调查外汇基金的刺激，在单细胞的生长条件，在2D，3D的器官在脊髓切片的细胞​​反应。在的脊髓cordslice是学习全国人大前体内的行为，移植后，因为在cytoarchitectonic的组织机构以及保存这些文化在9,10理想收件人组织。此外，这种体外模型还允许程序在技术上并不可行追踪细胞在体内使用在单细胞水平上的时间推移录影。这是极为必要的评估不仅是一个二维的环境中的细胞行为，而且在一个三维器官的条件， 在体内环境mimicks。该系统将允许使用玻璃盖的菜肴，在组织或器官培养， 在体外和体内的单个细胞迁移的3D跟踪，可以移动到前在vi中的中间步骤的高分辨率成像VO典范。

Watch this Scientific Journal Video about Electric Field-controlled Directed Migration of Neural Progenitor Cells in 2D and 3D Environments at JoVE.com

Electric Field-controlled Directed Migration of Neural Progenitor Cells in 2D and 3D Environments

此协议表明用于建立定制设计electrotactic的商会的二维和三维环境的方法，它可以追踪细胞<em&gt;在体内/体外</em使用时间推移录制在单细胞水平，以探讨趋电/电性和其他细胞的反应，直接电流（DC）电场（EFS）。

电场控制的二维和三维环境中的神经祖细胞定向迁移

Endogenous electric fields (EFs) occur naturally in vivo and play a critical role during tissue/organ development and regeneration, including that of the ...

Research

JoVE Journal

Bioengineering

Adaptation of a Haptic Robot in a 3T fMRI

触觉机器人适应在3T功能磁共振成像

Functional magnetic resonance imaging (fMRI) provides excellent functional brain imaging via the BOLD signal 1 with advantages including non-ionizing ...

科研

生物工程

Cultivation of Human Neural Progenitor Cells in a 3-dimensional Self-assembling Peptide Hydrogel

在三维自组装多肽水凝胶的人类神经祖细胞的培养

The  influence of 3-dimensional (3D) scaffolds on growth, proliferation and finally  neuronal differentiation is of great interest in order to find new ...

A Toolkit to Enable Hydrocarbon Conversion in Aqueous Environments

一个工具包，使水环境中的烃类转化

This work puts forward a toolkit that enables the conversion of alkanes by Escherichia coli and presents a proof of principle of its applicability. The ...

Ordering Single Cells and Single Embryos in 3D Confinement: A New Device for High Content Screening

订货单细胞和单胚胎的三维约束：高内涵筛选新设备

Biological cells are usually observed on flat (2D) surfaces. This condition is not physiological, and phenotypes and shapes are highly variable. Screening ...

Engineered 3D Silk-collagen-based Model of Polarized Neural Tissue

极化神经组织工程三维蚕丝胶原蛋白为基础的模式

Despite huge efforts to decipher the anatomy, composition and function of the brain, it remains the least understood organ of the human body. To gain a ...

Electrotaxis Studies of Lung Cancer Cells using a Multichannel Dual-electric-field Microfluidic Chip

使用多通道双电场微流控芯片肺癌细胞Electrotaxis研究

The behavior of directional cell migration under a direct current electric-field (dcEF) is referred to as electrotaxis. The significant role of ...

Improving 2D and 3D Skin In Vitro Models Using Macromolecular Crowding

Improving 2D and 3D Skin In Vitro Models Using Macromolecular Crowding

提高2D和3D皮肤<em&gt;体外</em&gt;模型使用大分子拥挤

The glycoprotein family of collagens represents the main structural proteins in the human body, and are key components of biomaterials used in modern ...

2D and 3D Matrices to Study Linear Invadosome Formation and Activity

2D和3D矩阵研究线性侵袭体形成和活性

Cell adhesion, migration, and invasion are involved in many physiological and pathological processes. For example, during metastasis formation, tumor ...

Characterizing Cell Migration Within Three-dimensional In Vitro Wound Environments

Characterizing Cell Migration Within Three-dimensional In Vitro Wound Environments

表征细胞迁移在三维体外伤口环境

Currently, most in vitro models of wound healing, such as well-established scratch assays, involve studying cell migration and wound closure on ...

Improved 3D Hydrogel Cultures of Primary Glial Cells for In Vitro Modelling of Neuroinflammation

Improved 3D Hydrogel Cultures of Primary Glial Cells for In Vitro Modelling of Neuroinflammation

改良的3D 水凝胶培养的原发胶质细胞的体外Neuroinflammation 模型的建立

In the central nervous system, numerous acute injuries and neurodegenerative disorders, as well as implanted devices or biomaterials engineered to enhance ...