Name: a galvanotaxis assay for analysis of neural precursor cell migration kinetics in an externally applied direct current electric field
Uploaded: 2012-10-13T14:00:00
Description: Watch this Scientific Journal Video about A Galvanotaxis Assay for Analysis of Neural Precursor Cell Migration Kinetics in an Externally Applied Direct Current Electric Field at JoVE.com

这项工作的经费由自然科学和工程研究理事会，加拿大（拨款＃249669，＃482986）和加拿大心脏及中风基金会（拨款＃485508）。作者感谢优素福埃尔 - 哈耶克和博士齐哇嗯为他们的协助开发实验的协议。

多伦多大学的动物保健委员会批准了在所有涉及动物处理的程序，按照制度指引（协议。20009387）。应当执行下面的方法，使用无菌的工具和技术，在层流罩（如适用）。 在下面的协议文本中，短语“EFH-SFM”是指以与表皮生长因子，碱性成纤维细胞生长因子和肝素的无血清培养基中。 EFH-SFM调查时，未分化的NPC，因为这些有丝分裂原的趋电维持其未分化状态的NPC 4。?...

<ol>
	<li>Borgens, R. B., Shi, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Uncoupling+histogenesis+from+morphogenesis+in+the+vertebrate+embryo+by+collapse+of+the+transneural+tube+potential.">Uncoupling histogenesis from morphogenesis in the vertebrate embryo by collapse of the transneural tube potential.</a> Dev. Dyn. 203, 456-467 (1995).</li><li>Song, B., Zhao, M., Forrester, J. V., McCaig, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrical+cues+regulate+the+orientation+and+frequency+of+cell+division+and+the+rate+of+wound+healing+in+vivo.">Electrical cues regulate the orientation and frequency of cell division and the rate of wound healing in vivo.</a> Proc. Natl. Acad. Sci. U.S.A. 99, 13577-13582 (2002).</li><li>Song, B., Zhao, M., Forrester, J., 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=Nerve+regeneration+and+wound+healing+are+stimulated+and+directed+by+an+endogenous+electrical+field+in+vivo.">Nerve regeneration and wound healing are stimulated and directed by an endogenous electrical field in vivo.</a> Journal of Cell Science. 117, 4681-4690 (2004).</li><li>Reynolds, B. A., Weiss, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Generation+of+neurons+and+astrocytes+from+isolated+cells+of+the+adult+mammalian+central+nervous+system.">Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system.</a> Science. 255, 1707-1710 (1992).</li><li>Reynolds, B. A., Weiss, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Clonal+and+population+analyses+demonstrate+that+an+EGF-responsive+mammalian+embryonic+CNS+precursor+is+a+stem+cell.">Clonal and population analyses demonstrate that an EGF-responsive mammalian embryonic CNS precursor is a stem cell.</a> Dev. Biol. 175, 1-13 (1996).</li><li>Chiasson, B. J., Tropepe, V., Morshead, C. M., vander Kooy, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adult+mammalian+forebrain+ependymal+and+subependymal+cells+demonstrate+proliferative+potential,+but+only+subependymal+cells+have+neural+stem+cell+characteristics.">Adult mammalian forebrain ependymal and subependymal cells demonstrate proliferative potential, but only subependymal cells have neural stem cell characteristics.</a> J. Neurosci. 19, 4462-4471 (1999).</li><li>Erickson, C. A., Nuccitelli, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Embryonic+fibroblast+motility+and+orientation+can+be+influenced+by+physiological+electric+fields.">Embryonic fibroblast motility and orientation can be influenced by physiological electric fields.</a> J. Cell Biol. 98, 296-307 (1984).</li><li>Zhao, M., Agius-Fernandez, A., Forrester, J. V., McCaig, C. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Orientation+and+directed+migration+of+cultured+corneal+epithelial+cells+in+small+electric+fields+are+serum+dependent.">Orientation and directed migration of cultured corneal epithelial cells in small electric fields are serum dependent.</a> J. Cell Sci. 109, 1405-1414 (1996).</li><li>Li, L. <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/Progenitor+Cell+Migration.">Direct-Current Electrical Field Guides Neuronal Stem/Progenitor Cell Migration.</a> Stem Cells. 26, 2193-2200 (2008).</li><li>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=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> Nat. Protoc. 2, 1479-1489 (2007).</li><li>Huang, C. -. W., Cheng, J. -. Y., Yen, M. -. H., Young, T. -. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Electrotaxis+of+lung+cancer+cells+in+a+multiple-electric-field+chip.">Electrotaxis of lung cancer cells in a multiple-electric-field chip.</a> Biosen Bioelectron. 24, 3510-3516 (2009).</li><li>SATO, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Input-output+relationship+in+galvanotactic+response+of+Dictyostelium+cells.">Input-output relationship in galvanotactic response of Dictyostelium cells.</a> Biosystems. 88, 261-272 (2007).</li><li>Meng, X. <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, 210-217 (2011).</li><li>Zhao, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Membrane+lipids,+EGF+receptors,+and+intracellular+signals+colocalize+and+are+polarized+in+epithelial+cells+moving+directionally+in+a+physiological+electric+field.">Membrane lipids, EGF receptors, and intracellular signals colocalize and are polarized in epithelial cells moving directionally in a physiological electric field.</a> The FASEB Journal. , (2002).</li><li>Fang, K., Ionides, E., Oster, G., Nuccitelli, R., Isseroff, R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidermal+growth+factor+receptor+relocalization+and+kinase+activity+are+necessary+for+directional+migration+of+keratinocytes+in+DC+electric+fields.">Epidermal growth factor receptor relocalization and kinase activity are necessary for directional migration of keratinocytes in DC electric fields.</a> J. Cell Sci. 112, 1967-1976 (1999).</li><li>McCaig, C. D. <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, 943-978 (2005).</li><li>Morshead, C. M., Benveniste, P., Iscove, N. N., vander Kooy, D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hematopoietic+competence+is+a+rare+property+of+neural+stem+cells+that+may+depend+on+genetic+and+epigenetic+alterations.">Hematopoietic competence is a rare property of neural stem cells that may depend on genetic and epigenetic alterations.</a> Nat. Med. 8, 268-273 (2002).</li><li>Babona-Pilipos, R., Droujinine, I. A., Popovic, M. R., Morshead, C. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adult+Subependymal+Neural+Precursors,+but+Not+Differentiated+Cells,+Undergo+Rapid+Cathodal+Migration+in+the+Presence+of+Direct+Current+Electric+Fields.">Adult Subependymal Neural Precursors, but Not Differentiated Cells, Undergo Rapid Cathodal Migration in the Presence of Direct Current Electric Fields.</a> PLoS ONE. 6, e23808 (2011).</li><li>Deuschl, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Randomized+Trial+of+Deep-Brain+Stimulation+for+Parkinson's+Disease.">A Randomized Trial of Deep-Brain Stimulation for Parkinson's Disease.</a> N. Engl. J. Med. 355, 896-908 (2006).</li><li>Stone, S. S. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stimulation+of+Entorhinal+Cortex+Promotes+Adult+Neurogenesis+and+Facilitates+Spatial+Memory.">Stimulation of Entorhinal Cortex Promotes Adult Neurogenesis and Facilitates Spatial Memory.</a> J. Neurosci. 31, 13469-13484 (2011).</li></ol>

该协议已经进行了调整，从以前的研究7-9的行之有效的方法。可以使用各种不同的技术，包括建造一个单独的玻璃以及禁闭细胞接种中部槽10,11的微细化，或使用CO 2激光烧蚀构造galvanotactic室。有些技术可能比别人更费力，代价高昂的。我们描述了一个简单和成本效益的分析，为构建全国人民代表大会趋电性室使用的材料通常存在于大多数细胞生物学实验室。我们的协议，...

<table border="1"><tbody><tr><td height="44"> 项目名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td></td><td></td><td></td><td> 神经前体细胞分离 </td></tr><tr><td> 2M的NaCl </td><td>西格玛</td><td> S5886 </td><td> 11.688克溶解在100毫升蒸馏水2 O </td></tr><tr><td> 1M氯化钾</td><td>西格玛</td><td> P5405 </td><td> 7.456克溶解在100毫升蒸馏水2 O </td></tr><tr><td> 1M MgCl 2的 </td><td>西格玛</td><td> M2393 </td><td> 20.33克溶解在100毫升蒸馏水2 O </td></tr><tr><td> 155毫米碳酸氢钠 </td><td>西格玛</td><td> S5761 </td><td> 1.302克溶解在100毫升蒸馏水2 O </td></tr><tr><td> 0.5M葡萄糖</td><td>西格玛</td><td> G6152 </td><td>溶解在1009.01克毫升蒸馏水2 O </td></tr><tr><td> 108毫米氯化钙 </td><td>西格玛</td><td> C7902 </td><td> 1.59克溶解在100毫升蒸馏水2 O </td></tr><tr><td>青霉素，链霉素</td><td> Gibco公司</td><td> 15070 </td><td></td></tr><tr><td>牛胰腺胰蛋白酶</td><td>西格玛</td><td> T1005 </td><td></td></tr><tr><td>羊睾丸透明质酸酶</td><td>西格玛</td><td> H6254 </td><td></td></tr><tr><td>犬尿喹啉酸</td><td>西格玛</td><td> K3375 </td><td></td></tr><tr><td>卵类粘蛋白胰蛋白酶抑制剂</td><td>沃辛顿</td><td> LS003086 </td><td></td></tr><tr><td> DMEM </td><td> Invitrogen公司</td><td> 12100046 </td><td></td></tr><tr><td> F12 </td><td> Invitrogen公司</td><td> 21700075 </td><td></td></tr><tr><td> 30％葡萄糖</td><td>西格玛</td><td> G6152 </td><td></td></tr><tr><td> 7.5％的NaHCO 3 </td><td>西格玛</td><td> S5761 </td><td></td></tr><tr><td> 1M HEPES </td><td>西格玛</td><td> H3375 </td><td> 23.83克溶解在100毫升蒸馏水2 O </td></tr><tr><td> L-谷氨酰胺</td><td> Gibco公司</td><td> 25030 </td><td></td></tr><tr><td> EGF </td><td> Invitrogen公司</td><td> PMG8041 </td><td>复溶在1毫升的激素组合和等分成20微升单位。 </td></tr><tr><td>成纤维细胞生长因子</td><td> Invitrogen公司</td><td> PHG0226 </td><td>复溶在0.5毫升的激素组合和等分成20微升单位。 </td></tr><tr><td>肝素</td><td>西格玛</td><td> H3149 </td><td></td></tr><tr><td> APO转</td><td> R＆D Systems公司</td><td> 3188-AT </td><td> 0.1 g溶于4毫升蒸馏水2 0 </td></tr><tr><td>腐胺</td><td>西格玛</td><td> P7505 </td><td>将9.61毫克到APO转，所以lution </td></tr><tr><td>胰岛素</td><td>西格玛</td><td> I5500 </td><td> 25毫克，溶于0.1N盐酸0.5毫升增加至3.5毫升卫生署2 0 </td></tr><tr><td>硒</td><td>西格玛</td><td> S9133 </td><td></td></tr><tr><td>黄体激素</td><td>西格玛</td><td> P6149 </td><td></td></tr><tr><td>标准的解剖工具</td><td>精细科学工具</td><td></td><td></td></tr><tr><td>解剖显微镜</td><td>蔡司</td><td> STEMI 2000 </td><td></td></tr><tr><td></td><td></td><td></td><td> 趋电池准备 </td></tr><tr><td>方形玻璃盖幻灯片</td><td> VWR </td><td> 16004 </td><td></td></tr><tr><td> 6N盐酸</td><td> VWR </td><td> BDH3204-1 </td><td></td></tr><tr><td>高真空润滑脂</td><td>道康宁</td><td></td><td></td></tr><tr><td> 60毫米陪替氏培养皿</td><td> Fisher Scientific则</td><td> 0875713A </td><td></td></tr><tr><td>聚-L-赖氨酸</td><td>西格玛</td><td> P4707 </td><td></td></tr><tr><td>基底膜</td><td> BD Biosciences公司</td><td> 354234 </td><td>解冻后取到150微升单位</td></tr><tr><td> FBS </td><td> Invitrogen公司</td><td> 10082139 </td><td>只有使用诱导NPC分化，以其他方式使用SFM + EFH文化传媒如上文所述的</td></tr><tr><td>计数显微镜</td><td>奥林巴斯</td><td> CKX41 </td><td></td></tr><tr><td></td><td></td><td></td><td> 活细胞延时成像 </td></tr><tr><td>银线</td><td>阿法埃莎</td><td> 11434 </td><td></td></tr><tr><td>超纯琼脂糖</td><td> Invitrogen公司</td><td> 15510-027 </td><td></td></tr><tr><td>热灭活ated FBS </td><td>西格玛</td><td> 16140071 </td><td></td></tr><tr><td> PVC管</td><td> Fisher Scientific则</td><td> 80000006 </td><td> 3/32“ID×5/32”OD </td></tr><tr><td>漂白</td><td>高乐氏（Clorox） </td><td></td><td></td></tr><tr><td> 10毫升注射器</td><td> BD </td><td> 309604 </td><td></td></tr><tr><td> 18号针</td><td> BD </td><td> 305195 </td><td></td></tr><tr><td> DREMEL钻</td><td> DREMEL </td><td>型号750 </td><td></td></tr><tr><td>倒置显微镜配备了加湿，孵化室</td><td>蔡司</td><td> Axiovert-200M </td><td></td></tr><tr><td></td><td></td><td></td><td>食谱<table border="1"><tbody><tr><td> 项目 </td><td> 量 </td></tr><tr><td> 2M的NaCl </td><td> 6.2毫升</td></tr><tr><td>1M氯化钾</td><td> 0.5毫升</td></tr><tr><td> 1M MgCl 2的 </td><td> 0.32毫升</td></tr><tr><td> 155MM 碳酸氢钠 </td><td> 16.9毫升</td></tr><tr><td> 1M葡萄糖</td><td> 1毫升</td></tr><tr><td> 108毫米氯化钙 </td><td> 0.09256毫升</td></tr><tr><td>青霉素，链霉素</td><td> 1毫升</td></tr><tr><td>无菌水</td><td> 74毫升</td></tr></tbody></table> 人工脑脊液 <table border="1"><tbody><tr><td> 项目 </td><td> 体积或质量 </td></tr><tr><td>人工脑脊液</td><td> 30毫升</td></tr><tr><td>牛胰腺胰蛋白酶</td><td> 40毫克</td></tr><tr><td>羊睾丸透明质酸酶</td><td> 22.8毫克</td></tr><tr><td>犬尿喹啉酸</td><td> 5毫克</td></tr></tbody></table> &lt;STRONG&gt;胰蛋白酶溶液<table border="1"><tbody><tr><td> 项目 </td><td> 体积或质量 </td></tr><tr><td> SFM </td><td> 15毫升</td></tr><tr><td>卵类粘蛋白胰蛋白酶抑制剂</td><td> 10毫克</td></tr></tbody></table> 胰蛋白酶抑制剂解决方案 <table border="1"><tbody><tr><td> 项目 </td><td> 量 </td></tr><tr><td>无菌水</td><td> 37毫升</td></tr><tr><td> 10X DMEM/F12 </td><td> 10毫升</td></tr><tr><td> 30％葡萄糖</td><td> 2毫升</td></tr><tr><td> 7.5％的NaHCO 3 </td><td> 1.5毫升</td></tr><tr><td> 1M HEPES </td><td> 0.5毫升</td></tr><tr><td>转铁蛋白，腐胺溶液</td><td> 4毫升</td></tr><tr><td> 25毫克胰岛素的解决方案</td><td> 4毫升</td></tr><tr><td>硒</td><td> 100亩; L </td></tr><tr><td>黄体激素</td><td> 100微升</td></tr></tbody></table> 激素组合 （100毫升总，储存在-20°C） <table border="1"><tbody><tr><td> 项目 </td><td> 量 </td></tr><tr><td>无菌水</td><td> 37.5毫升</td></tr><tr><td> 10X的DMEM/F12（3:1） </td><td> 5毫升</td></tr><tr><td> 30％葡萄糖</td><td> 1毫升</td></tr><tr><td> 7.5％的NaHCO 3 </td><td> 0.75毫升</td></tr><tr><td> 1M HEPES </td><td> 0.25毫升</td></tr><tr><td>激素组合</td><td> 5毫升</td></tr><tr><td> L-谷氨酰胺</td><td> 0.5毫升</td></tr><tr><td>青霉素，链霉素</td><td> 0.5毫升</td></tr></tbody></table> EFH-SFM无血清培养基：添加10μl的表皮生长因子，10μl的成纤维细胞生长因子，肝素FBS-SFM和3.66微升：添加0.5毫升FBS <ta表边境=“1”&gt; <tbody><tr><td> 项目 </td><td> 量 </td></tr><tr><td>基底膜</td><td> 150微升</td></tr><tr><td> SFM </td><td> 3.6毫升</td></tr></tbody></table>应Matrigel的解决方案 Matrigel的等分试样放置在一个盒子里的冰和允许缓慢解冻超过4-5小时，以形成一种粘稠的液体混合之前与SFM。这将确保形成一层光滑的Matrigel基板。如果不解冻缓慢，导致基板包含一丛丛的Matrigel，可能会阻碍细胞的迁移。 <table border="1"><tbody><tr><td> 项目 </td><td> 体积或质量 </td></tr><tr><td>超纯琼脂糖</td><td>在10毫升DDH 2 O 300毫克</td></tr><tr><td> SFM 热灭活FBS </td><td> 8毫升 2毫升</td></tr></tbody></table> 基底膜解决方案MIX 8毫升SFM 2毫升热灭活FBS在15毫升猎鹰管。混合琼脂糖用10ml DDH 2 0的埃伦迈尔烧瓶中，并在10秒的间隔在30秒的微波的热量，以确保从微波炉中取出该溶液后，每10秒时间间隔，并彻底混合。最后10秒的微波期，混合的的琼脂糖溶液与SFM / FBS的解决方案和存储在57℃水浴。 </td></tr></tbody></table>

a galvanotaxis assay for analysis of neural precursor cell migration kinetics in an externally applied direct current electric field

神经干细胞和祖细胞（统称为神经前体细胞）（的NPC）在成年哺乳动物大脑的发现已导致一个机构的研究，目的是在利用这些细胞的多能和增殖特性的发展的神经再生策略。这种战略成功的一个关键步骤是动员的NPC外源性移植后向病变部位或提高内源性的前体，发现脑室周围白质区域的中枢神经系统的反应。因此，有必要了解的机制，促进，引导，增强人大迁移。我们的工作主要集中在利用直流电场（量增多），以促进和直接的NPC迁移 - 这种现象被称为趋电性。内源性生理电场的正常发育和伤口修复细胞迁移过程中发挥作用的关键线索。药理中断在蝾螈胚胎的的跨神经管潜力会导致严重的发育畸形。的速度修复受伤的角膜伤口愈合的情况下，直接与上皮损伤后产生的伤害潜力的幅度，药理增强或中断，这DCEF 2-3所示。我们已经证明，成年室管膜下的NPC进行快速和导演的阴极迁移的影响 ，当暴露在外部施加的dcEF。在这个协议中，我们将介绍创建一个简单而有效的高分辨率，长期观察的细胞体易位（迁移）在单细胞水平的趋电检测实验室的技术。该测定中，将适合于调查机制调节dcEF转导到细胞运动通过使用转基因或基因敲除小鼠，短干扰RNA，或特定的受体激动剂/拮抗剂。

运动学分析表明，在250毫伏/毫米dcEF的存在下，未分化的NPC表现出高度定向和快速的趋电向阴极（ 图5A，电影1）。在一个dcEF缺席，随机运动的细胞中观察到（ 图5B，电影2）。在此输入强度，&gt; 98％的未分化的NPC迁移整个6-8小时，这是他们的成像，并自死细胞不迁移，这表明，它们在此期间，在存在或不存在的dcEF仍然可行。 有区别的表型进行可以忽...

Watch this Scientific Journal Video about A Galvanotaxis Assay for Analysis of Neural Precursor Cell Migration Kinetics in an Externally Applied Direct Current Electric Field at JoVE.com

A Galvanotaxis Assay for Analysis of Neural Precursor Cell Migration Kinetics in an Externally Applied Direct Current Electric Field

在这个协议中，我们将演示如何构建自定义室，允许应用程序的直流电场，使时间的推移成像的成人脑源性神经前体细胞转趋电性。

一个的趋电含量为神经前体细胞迁移的动力学分析外部施加直流电场

The discovery of neural stem and progenitor  cells (collectively termed neural precursor cells) (NPCs) in the adult  mammalian brain has led to a body of ...

Research

JoVE Journal

Neuroscience

Neural-Colony Forming Cell Assay: An Assay To Discriminate Bona Fide Neural Stem Cells from Neural Progenitor Cells

神经细胞集落形成细胞分析检测，以区分善意的神经祖细胞的神经干细胞

The neurosphere assay (NSA) is one of the most frequently used methods to isolate, expand and also calculate the frequency of neural stem cells (NSCs). ...

科研

神经科学

Electrode Positioning and Montage in Transcranial Direct Current Stimulation

经颅直接电流刺激电极定位和蒙太奇

Transcranial direct current stimulation (tDCS) is a technique that has been intensively investigated in the past decade as this method offers a ...

Analysis of Trunk Neural Crest Cell Migration using a Modified Zigmond Chamber Assay

躯干神经嵴细胞迁移使用一个修改过的Zigmond室法分析

Neural crest cells (NCCs) are a transient population of cells present in vertebrate development that emigrate from the dorsal neural tube (NT) after ...

Reproducible Mouse Sciatic Nerve Crush and Subsequent Assessment of Regeneration by Whole Mount Muscle Analysis

重复性小鼠坐骨神经挤压和再生的后续评估整装肌肉分析

Regeneration in the peripheral nervous system (PNS) is widely studied both for its relevance to human disease and to understand the robust regenerative ...

Analysis of Neural Crest Migration and Differentiation by Cross-species Transplantation

跨物种移植神经嵴迁移和分化的分析

Avian embryos provide a unique platform for studying many vertebrate developmental processes, due to the easy access of the embryos within the egg. ...

The Neuroblast Assay: An Assay for the Generation and Enrichment of Neuronal Progenitor Cells from Differentiating Neural Stem Cell Progeny Using Flow Cytometry

神经母细胞含量：从区分神经干细胞后代使用流式细胞仪，神经祖细胞的生成和富集含量

Neural stem cells (NSCs) can be isolated and expanded in large-scale, using the neurosphere assay and differentiated into the three major cell types of ...

Isolation and Culture of Neural Crest Cells from Embryonic Murine Neural Tube

从小鼠胚胎神经管的神经嵴细胞的分离和培养

The embryonic neural crest (NC) is a multipotent progenitor population that originates at the dorsal aspect of the neural tube, undergoes an epithelial to ...

A Quantitative Cell Migration Assay for Murine Enteric Neural Progenitors

定量细胞迁移检测方法的鼠肠神经前体细胞

Neural crest cells (NCC) are a transient and multipotent cell population that originates from the dorsal neural tube and migrates extensively throughout ...

Long-term Behavioral Tracking of Freely Swimming Weakly Electric Fish

自由游泳弱电鱼的长期行为跟踪

Long-term behavioral tracking can capture and quantify natural animal behaviors, including those occurring infrequently. Behaviors such as exploration and ...

Remotely Supervised Transcranial Direct Current Stimulation: An Update on Safety and Tolerability

远程监督的经颅直流电刺激: 安全性和耐受的最新进展

The remotely supervised tDCS (RS-tDCS) protocol enables participation from home through guided and monitored self-administration of tDCS treatment while ...