Name: migration, chemo-attraction, and co-culture assays for human stem cell-derived endothelial cells and gabaergic neurons
Uploaded: 2020-01-23T12:00:00
Description: Watch this Scientific Journal Video about Migration, Chemo-Attraction, and Co-Culture Assays for Human Stem Cell-Derived Endothelial Cells and GABAergic Neurons at JoVE.com

这项工作得到了国家精神卫生研究所（R01MH110438）和国家神经疾病和中风研究所（R01NS100808）颁发的AV奖的支持。

1. 人类前皮内皮细胞的培养与储存
<ol><li>在地下膜基质涂层（见材料表）6孔板（E6培养基包含50纳克/mL VEGF-A、100纳克/mL FGF2和5μM GABA）的表）中保持人体腹膜内皮细胞，温度为37°C和5%CO2。每隔一天更换媒体。</li>
	<li>在4°C下解冻基底膜基质，在冷DMEM/F12介质中稀释，使其溶液达到1：100。用1 mL的基质溶液涂抹每口6孔板。在37°C下孵育板至少1小时，然后再使用。</li>
	<li>允许人...

<ol>
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L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Induced+pluripotent+stem+cells:+the+new+patient?.">Induced pluripotent stem cells: the new patient?.</a> Nature Reviews Molecular Cell Biology. 13 (11), 713-726 (2012).</li><li>Ardhanareeswaran, K., Mariani, J., Coppola, G., Abyzov, A., Vaccarino, F. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Human+induced+pluripotent+stem+cells+for+modelling+neurodevelopmental+disorders.">Human induced pluripotent stem cells for modelling neurodevelopmental disorders.</a> Nature Reviews Neurology. 13 (5), 265-278 (2017).</li><li>Stubbs, D., 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=Neurovascular+congruence+during+cerebral+cortical+development.">Neurovascular congruence during cerebral cortical development.</a> Cerebral Cortex. 19 (1), 32-41 (2009).</li><li>Vissapragada, R., 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=Bidirectional+crosstalk+between+periventricular+endothelial+cells+and+neural+progenitor+cells+promotes+the+formation+of+a+neurovascular+unit.">Bidirectional crosstalk between periventricular endothelial cells and neural progenitor cells promotes the formation of a neurovascular unit.</a> Brain Research. 1565, 8-17 (2014).</li><li>JoVE Science Education Database. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cell+Biology.+The+Transwell+Migration+Assay.">Cell Biology. The Transwell Migration Assay.</a> Journal of Visualized Experiments. , (2019).</li><li>Renaud, J., Martinovic, M. G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Development+of+an+insert+co-culture+system+of+two+cellular+types+in+the+absence+of+cell-cell+contact.">Development of an insert co-culture system of two cellular types in the absence of cell-cell contact.</a> Journal of Visualized Experiments. 113, e54356 (2016).</li><li>Guan, J. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In+vitro+scratch+assay:+a+convenient+and+inexpensive+method+for+analysis+of+cell+migration+in+vitro.">In vitro scratch assay: a convenient and inexpensive method for analysis of cell migration in vitro.</a> Nature Protocols. 2 (2), 329-333 (2007).</li><li>Nelson, R. D., Quie, P. G., Simmons, R. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Chemotaxis+under+agarose:+a+new+and+simple+method+for+measuring+chemotaxis+and+spontaneous+migration+of+human+polymorphonuclear+leukocytes+and+monocytes.">Chemotaxis under agarose: a new and simple method for measuring chemotaxis and spontaneous migration of human polymorphonuclear leukocytes and monocytes.</a> The Journal of Immunology. 115 (6), 1650-1656 (1975).</li><li>Zigmond, S. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Ability+of+polymorphonuclear+leukocytes+to+orient+in+gradients+of+chemotactic+factors.">Ability of polymorphonuclear leukocytes to orient in gradients of chemotactic factors.</a> Journal of Cell Biology. 75 (2), 606-616 (1977).</li><li>Zicha, D., Dunn, G., Jones, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Analyzing+chemotaxis+using+the+Dunn+direct-viewing+chamber.">Analyzing chemotaxis using the Dunn direct-viewing chamber.</a> Methods in Molecular Biology. 75, 449-457 (1997).</li><li>Kim, B. J., Wu, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Microfluidics+for+mammalian+cell+chemotaxis.">Microfluidics for mammalian cell chemotaxis.</a> Annals of Biomedical Engineering. 40 (6), 1316-1327 (2012).</li></ol>

在这里，我们描述了三个体外测定，一起提供定量评估的人类腹膜内皮细胞特异性特性。这些测定对于获得人类外皮内皮细胞与人类内皮细胞相互作用的机械性见解将非常有价值。使用配体、抑制剂或具有基因特异性敲除或过度表达的细胞的实验将识别或验证介导内皮细胞引导的内神经元迁移或腹膜内皮细胞远距离迁移特性的分子。这些测定还可以修改以执行活细胞延时迁移研究。此外，还有证?...

内皮细胞形成血管的衬里，调节重要功能，包括维持血管壁渗透性、调节血流、血小板聚集和形成新血管。在大脑中，内皮细胞构成一个关键的血液-脑屏障的一部分，它严格控制大脑和血液之间的物质交换。我们在过去十年的研究已经确定了大脑内皮细胞的神经原发生新作用，这些作用对大脑发育和行为2、3、4、5有重要影响。我们已经表明，小鼠胚胎前脑是由两个不同的子类型的血管，皮面和腹周血管，在解剖学，起源和发育轮廓2不同。内皮细胞衬里这两个血管亚型显示其基因表达配置文件明显的差异。虽然皮叶细胞大多表达与炎症和免疫反应相关的基因，但腹腔内皮细胞在表达通常与神经发生、神经元迁移、化学和轴神指导3相关的基因时，具有独特的丰富性。外皮内皮细胞也拥有一种新的GABA信号通路，不同于传统的神经元GABA信号通路5。与其基因表达相伴，发现外膜内皮细胞调节发育中新皮质中GABAergic内神经元的迁移和分布。在胚胎发育过程中，腹腔内皮细胞沿腹膜-背梯度进行远距离迁移，以建立腹腔血管网络2、3。一天后，内神经内向反映了这种迁徙路线。迁移的内神经元与预先形成的外皮血管网络进行物理交互，并用作在新皮质到达最终目的地的指南。除了作为物理基质外，腹膜内皮细胞还充当迁移神经元的导航线索来源。前心内皮细胞分泌GABA指导内神经元迁移并调节其最终分布模式4。内神经元迁移和分布的缺陷与神经精神病有关，如自闭症、癫痫、精神分裂症和抑郁症6，7，8，9，10。因此，研究前心室内皮细胞功能及其对人类环境中的内神经元迁移的影响，对于解决这些疾病的发病机制至关重要。
我们利用诱导多能干细胞（iPSC）技术12、13，从人类胚胎干细胞中生成了人类造血干细胞。为了验证人类腹膜内皮细胞是否忠实地模仿小鼠腹膜内皮细胞，并定量评估它们对内神经元迁移的影响，我们开发了三种体外测定：远距离迁移测定、共培养迁移测定和化学吸引力测定。在这里，我们详细描述了这些检测的协议。所有三种检测都基于硅培养插件的使用，以创建一个小矩形的细胞补丁（固定尺寸），周围环绕着无细胞空间。迁移距离是通过测量单元格的最终位置与第 0 天已勾勒的矩形贴片边框之间的距离来评估的。在远距离迁移测定中，人类腹膜内皮细胞在35毫米盘的中心作为斑块播种，并计算细胞在长时间内所经过的距离。在共培养迁移测定中，人类腹膜内皮细胞与人类内膜细胞共同播种，作为35毫米盘中的一个斑块。此设置允许检查这两种细胞类型的直接物理相互作用对内神经元迁移速率的影响。化学吸引力测定测量内神经元的迁移，以响应人类腹膜内皮细胞分泌的化学吸引力线索。内侧神经元被播种为矩形斑块，与人腹膜内皮细胞和控制非外膜内皮细胞播种作为类似大小的补丁在两侧。每个细胞斑块都通过500μm的无细胞间隙分离。 与对照非外皮内皮细胞相比，通过量化向外皮内皮细胞迁移的细胞数量来评估内神经元的反应。
这些测定提供了对人腹膜内皮细胞功能及其对内神经元迁移的影响的有力评估。长距离测定和共培养迁移测定的新颖设置提供了厘米（±1-1.5 cm）范围内的无细胞空间，允许远距离迁移检测。表1概述了我们与其他常用检测相比的检测特征。总体而言，这里描述的测定将作为一个平台，用于评估"患病"的外室内皮细胞和由精神分裂症、自闭症或癫痫等脑疾病的iPSC产生的内神经元。这些测定也可用于确定不同条件（如抑制剂、配体、RNAi）如何影响细胞迁移。最后，这些测定可以针对其他细胞类型进行优化，以测量远距离迁移、化学吸引或细胞介导迁移。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>Accutase dissociation solution</td>
			<td>Millipore Sigma</td>
			<td>SCR005</td>
			<td>Cell dissociation solution (for periventricular endothelial cells, step 1.4)</td>
		</tr>
		<tr>
			<td>Anti-human &#946;-Tubulin antibody</td>
			<td>Biolegend</td>
			<td>802001</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-human CD31 antibody</td>
			<td>Millipore Sigma</td>
			<td>CBL468</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti- MAP2 antibody</td>
			<td>Neuromics</td>
			<td>CH22103</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-active Caspase 3 antibody</td>
			<td>Millipore Sigma</td>
			<td>AB3623</td>
			<td></td>
		</tr>
		<tr>
			<td>Control human endothelial cells</td>
			<td>Cellular Dynamics</td>
			<td>R1022</td>
			<td></td>
		</tr>
		<tr>
			<td>Control endothelial Cells Medium Supplement</td>
			<td>Cellular Dynamics</td>
			<td>M1019</td>
			<td></td>
		</tr>
		<tr>
			<td>Cryogenic vials</td>
			<td>Fisher Scientific</td>
			<td>03-337-7Y</td>
			<td></td>
		</tr>
		<tr>
			<td>DMEMF/12 medium</td>
			<td>Thermofisher Scientific</td>
			<td>11320033</td>
			<td></td>
		</tr>
		<tr>
			<td>DMSO</td>
			<td>Sigma-Aldrich</td>
			<td>D2650</td>
			<td></td>
		</tr>
		<tr>
			<td>E6 medium</td>
			<td>Thermofisher Scientific</td>
			<td>A1516401</td>
			<td></td>
		</tr>
		<tr>
			<td>FGF2</td>
			<td>Thermofisher Scientific</td>
			<td>PHG0261</td>
			<td></td>
		</tr>
		<tr>
			<td>Fibronectin</td>
			<td>Thermofisher Scientific</td>
			<td>33016-015</td>
			<td></td>
		</tr>
		<tr>
			<td>Freezing Container</td>
			<td>Thermofisher Scientific</td>
			<td>5100</td>
			<td></td>
		</tr>
		<tr>
			<td>GABA</td>
			<td>Sigma-Aldrich</td>
			<td>A2129</td>
			<td></td>
		</tr>
		<tr>
			<td>Hemacytometer</td>
			<td>Sigma-Aldrich</td>
			<td>Z359629</td>
			<td></td>
		</tr>
		<tr>
			<td>Human GABAergic neurons</td>
			<td>Cellular Dynamics</td>
			<td>R1013</td>
			<td></td>
		</tr>
		<tr>
			<td>Human GABAergic neurons base medium</td>
			<td>Cellular Dynamics</td>
			<td>M1010</td>
			<td></td>
		</tr>
		<tr>
			<td>Human GABAergic neuron Neural supplement</td>
			<td>Cellular Dynamics</td>
			<td>M1032</td>
			<td></td>
		</tr>
		<tr>
			<td>Laminin</td>
			<td>Sigma</td>
			<td>L2020</td>
			<td></td>
		</tr>
		<tr>
			<td>Matrigel</td>
			<td>Corning</td>
			<td>356230</td>
			<td>Basement membrane matrix</td>
		</tr>
		<tr>
			<td>Mounting Medium</td>
			<td>Vector laboratories</td>
			<td>H-1200</td>
			<td></td>
		</tr>
		<tr>
			<td>poly-L-ornithin</td>
			<td>Sigma</td>
			<td>p4957</td>
			<td></td>
		</tr>
		<tr>
			<td>PBS</td>
			<td>Thermofisher Scientific</td>
			<td>14190</td>
			<td></td>
		</tr>
		<tr>
			<td>Trypan blue</td>
			<td>Thermofisher Scientific</td>
			<td>15250061</td>
			<td></td>
		</tr>
		<tr>
			<td>TrypLE</td>
			<td>Thermofisher Scientific</td>
			<td>12563011</td>
			<td>Cell dissociation solution (for GABAergic interneurons and endothelial cells, sections 3 and 4)</td>
		</tr>
		<tr>
			<td>VEGF-A</td>
			<td>Peprotech</td>
			<td>100-20</td>
			<td></td>
		</tr>
		<tr>
			<td>VascuLife VEGF Medium Complete Kit</td>
			<td>Lifeline Cell Technologies</td>
			<td>LL-0003</td>
			<td>Component of control human endothelial cell medium</td>
		</tr>
		<tr>
			<td>2-well silicone culture-Insert</td>
			<td>ibidi</td>
			<td>80209</td>
			<td></td>
		</tr>
		<tr>
			<td>3-well silicone culture-Insert</td>
			<td>ibidi</td>
			<td>80369</td>
			<td></td>
		</tr>
		<tr>
			<td>35 mm dish</td>
			<td>Corning</td>
			<td>430165</td>
			<td></td>
		</tr>
		<tr>
			<td>15-ml conical tube</td>
			<td>Fisher Scientific</td>
			<td>07-200-886</td>
			<td></td>
		</tr>
		<tr>
			<td>4% PFA solution</td>
			<td>Fisher Scientific</td>
			<td>AAJ19943K2</td>
			<td></td>
		</tr>
		<tr>
			<td>6-well tissue culture plate</td>
			<td>Fisher Scientific</td>
			<td>14-832-11</td>
			<td></td>
		</tr>
		<tr>
			<td>Inverted phase contrast microscope</td>
			<td>Zeiss</td>
			<td>Zeiss Axiovert 40C</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluorescent microscope</td>
			<td>Olympus</td>
			<td>FSX-100</td>
			<td></td>
		</tr>
	</tbody>
</table>

migration, chemo-attraction, and co-culture assays for human stem cell-derived endothelial cells and gabaergic neurons

脑血管在神经系统发育和脑病病因中的作用日益受到重视。我们最近的研究已经确定了一种特殊的血管细胞群体，即腹膜内皮细胞，在胚胎发育期间前脑GABAergic内神经元的迁移和分布中起着关键作用。这，加上他们的细胞自主功能，暗示了前心室内皮细胞在神经精神病（如精神分裂症、癫痫和自闭症）的病理学中的新作用。在这里，我们描述了三种不同的体外测定，它们共同评估腹膜内皮细胞的功能及其与GABAergic内神经元的相互作用。使用这些检测方法，特别是在人类环境中，将使我们能够识别腹膜内皮细胞和大脑疾病之间的联系。这些测定简单、成本低、可重复，可轻松适应任何粘附细胞类型。

在 35 mm 的培养皿内设置单井培养插件的步骤如图1所示。远距离迁移测定和共培养迁移测定使用单井插入物在聚L-Ornithine/拉米宁涂层35毫米盘的中心播种所需数量的细胞。第 0 天，单元格以矩形面片形式存在（图 2A，C）。在第 0 天图像中，可通过像元图层的锐边（图 2C中的白色虚?...

Watch this Scientific Journal Video about Migration, Chemo-Attraction, and Co-Culture Assays for Human Stem Cell-Derived Endothelial Cells and GABAergic Neurons at JoVE.com

Migration, Chemo-Attraction, and Co-Culture Assays for Human Stem Cell-Derived Endothelial Cells and GABAergic Neurons

我们提出了三个简单的体外测定-远距离迁移测定，共培养迁移测定，和化学吸引力测定-共同评估人类干细胞衍生的腹膜内皮细胞的功能及其与GABAergic内神经的相互作用。

人类干细胞衍生内皮细胞和GABAergic神经元的迁移、化学吸引和共培养测定

Role of brain vasculature in nervous system development and etiology of brain disorders is increasingly gaining attention. Our recent studies have ...

该协议描述了三种体外分析，这些测定共同评估了人类近端细胞的功能及其与人类GABAergic内创细胞的相互作用。这些测定的结果将提供对内皮细胞和脑部疾病之间联系的关键洞察。这些测定简单、低成本，允许在厘米范围内测量细胞迁移，而其他现有测定无法实现。GABAergic内科患者的迁移和分布缺陷与精神疾病有关，如自闭症、癫痫、精神分裂症和抑郁症。因此，研究在人类环境中的透镜内皮细胞与GABAergic内皮细胞的相互作用对于解决这些疾病的发病机制至关重要。首先为测定准备人体细胞。允许人类外皮细胞达到70%至80%的汇合，然后根据手稿的指示分离它们，并使用 trypan 蓝色排除法计算它们。准备人类GABAergic内科龙，温暖的细胞分离溶液和在37摄氏度的神经元介质等分10分钟。然后从每个含有细胞的井中吸出介质，然后用每井一毫米的 PBS 清洗。通过每井添加0.5毫升预谋的分离溶液，并在37摄氏度下孵育5分钟，分离细胞。孵育后，每井加入一毫升神经元介质，将细胞溶液转移到15毫升锥形管中，轻轻三角分离细胞团。在室温下以380倍G离心细胞5分钟，吸升液，将细胞颗粒重新在一毫升神经元培养中。然后使用 trypan 蓝色排除数活细胞。在使用前10分钟，在37摄氏度的温度下加热细胞分离溶液和内皮介质的等分，准备控制人类内皮细胞。从每一个井中吸出介质，用每井一毫升的PBS清洗。将预预热分离溶液的0.5毫升添加到每井中，并在室温下孵育板5分钟，从而分离细胞。然后，每井加入一毫升内皮细胞培养，以中和分离溶液，并将细胞转移到15毫升锥形管中。将细胞在200倍G下离心5分钟，吸升液，并在一毫升内皮细胞培养中重新分配细胞。然后使用 trypan 排除方法对单元格进行计数。使用无菌刀片切割两井硅胶培养件的三面，准备一个井培养物刀片。用无菌钳子取出刀片，并将其放在聚 L-ornithine 和层压涂层盘的中心，然后沿着刀片的边缘按压以将其固定到表面上。小心地将盘子倒置，以验证刀片是否牢固地粘附，并使用永久黑市用超细尖端标记刀片室的边界。将人体GABAergic内膜在神经元培养基和人类外向性内皮细胞和内皮细胞培养基中悬浮，浓度为每70微升3万，然后在每个井培养物插入体内播种70微升细胞溶液。在神经元培养皿中加入一毫升神经元培养基，以填充刀片周围的区域，防止涂层干燥。同样，在心室内皮细胞培养皿中加入一毫升内皮细胞介质。在显微镜下检查细胞，以验证它们是否从插入室泄漏，然后在37摄氏度和5%的二氧化碳下孵育24小时。孵育后，在显微镜下再次检查细胞，以验证它们是否正确连接。播种48小时后，用无菌钳子轻轻取出刀片，并在显微镜下检查细胞，以验证细胞层是否不受干扰。从神经元和外皮细胞培养皿中去除培养基，并给每个菜中加入一毫升新鲜培养基。在37摄氏度和5%的二氧化碳下孵育细胞5天。然后，取出介质，用4%PFA固定细胞10分钟，然后用PBS洗涤三次。为了进行共培养测定，在共培养培养基的70微升中悬浮3万个GABAergic内膜和3万个人类透镜内皮细胞，然后在一个井插入舱内播种这种细胞溶液。48 小时后，拆下刀片。在37摄氏度和5%的二氧化碳下孵育5天。孵育后，取出介质，用4%PFA固定细胞，用PBS洗涤三次。要执行化学吸引力测定，在聚 L-ornithine 和层压涂层的 35 毫米盘的中心放置三井培养物，将培养皿倒置，并使用带超细尖端的永久标记标记刀片的中间隔间周围的边界。种子3万GABAergic内皮瘤在70微升的神经元介质在中间的隔间，然后种子10，000个透体内皮细胞和10，000控制内皮细胞和70微升各自的介质在两个外层隔间。沿菜的侧面加入一毫升共培养基，以防止盘子上的涂层干燥。48小时后，取出插入物，在37摄氏度和5%的二氧化碳下孵育细胞36小时。孵育后，取出介质，固定细胞，用PBS洗涤三次。长距离和共培养迁移分析用于研究心室内皮细胞和GABAergic神经元之间的相互作用。在长距离迁移测定中，细胞在第 0 天开始作为矩形斑块，但到 48 小时后，它们已迁移到无细胞空间。具有抗活性卡斯帕塞-3抗体的免疫细胞化学染色，是凋亡的标记物，在种子神经元中未显示凋亡信号。在联合培养迁移测定中，当内维龙与人类外向性内皮细胞共同播种时，神经元与单独播种或与对照内皮细胞共同播种时相比，距离更远。在化疗吸引力测定中，与对照性内皮细胞相比，向内皮内皮细胞迁移的内皮细胞数量显著增加，这证实了GABAergic内皮龙对由透卵管内皮细胞分泌的化疗吸引力线索有选择性的反应。这些测定可以使用配体、抑制剂或分析法进行修饰，从而获得对人类外向性内皮细胞与人类内皮素相互作用的机械见解。它们还可用于研究人类外皮细胞与其他神经细胞的相互作用，如我们小组和其他人报告。尝试此过程时，必须将刀片牢固地固定在培养皿上，并在整个检测过程中保持不受干扰，否则可能导致插入物与绘制边界发生细胞泄漏、细胞分离或错位。我们的工作已经表明，人类近端膜细胞在精神分裂症、癫痫和自闭症等精神疾病的发病机制中具有细胞自主作用。因此，这些检测将允许使用IPSC技术评估这些疾病患者衍生的病性内皮细胞。

Research

JoVE Journal

Neuroscience

Bilaminar Co-culture of Primary Rat Cortical Neurons and Glia

二层共培养小学的大鼠皮层神经元和神经胶质

This video will guide you through the process of culturing rat cortical neurons in the presence of a glial feeder layer, a system known as a bilaminar or ...

科研

神经科学

Inhibitory Synapse Formation in a Co-culture Model Incorporating GABAergic Medium Spiny Neurons and HEK293 Cells Stably Expressing GABAA Receptors

Inhibitory Synapse Formation in a Co-culture Model Incorporating GABAergic Medium Spiny Neurons and HEK293 Cells Stably Expressing GABAA Receptors

在共同的文化整合模式中GABA能神经元的刺和HEK293细胞抑制性突触形成稳定表达GABA<sub&gt;一个</sub&gt;受体

Inhibitory neurons act in the central nervous system to regulate the dynamics and spatio-temporal co-ordination of neuronal networks. GABA ...

Dorsal Root Ganglia Neurons and Differentiated Adipose-derived Stem Cells: An In Vitro Co-culture Model to Study Peripheral Nerve Regeneration

Dorsal Root Ganglia Neurons and Differentiated Adipose-derived Stem Cells: An In Vitro Co-culture Model to Study Peripheral Nerve Regeneration

背根神经节神经元和区分脂肪干细胞：一种<em&gt;在体外</em&gt;共培养模型来研究周围神经再生

Dorsal root ganglia (DRG) neurons, located in the intervertebral foramina of the spinal column, can be used to create an in vitro system facilitating the ...

Enumeration of Neural Stem Cells Using Clonal Assays

使用克隆测定神经干细胞的计数。

Neural stem cells (NSCs) have the ability to self-renew and generate the three major neural lineages &#8212; astrocytes, neurons and oligodendrocytes. ...

Modelling Zika Virus Infection of the Developing Human Brain In Vitro Using Stem Cell Derived Cerebral Organoids

Modelling Zika Virus Infection of the Developing Human Brain In Vitro Using Stem Cell Derived Cerebral Organoids

利用干细胞衍生脑 Organoids体外培养人脑组织 Zika 病毒感染的模拟研究

The recent emergence of Zika virus (ZIKV) in susceptible populations has led to an abrupt increase in microcephaly and other neurodevelopmental conditions ...

Method for High Speed Stretch Injury of Human Induced Pluripotent Stem Cell-derived Neurons in a 96-well Format

96井格式人诱导多潜能干细胞源神经元高速拉伸损伤的方法

Traumatic brain injury (TBI) is a major clinical challenge with high morbidity and mortality. Despite decades of pre-clinical research, no proven ...

Compartmentalization of Human Stem Cell-Derived Neurons within Pre-Assembled Plastic Microfluidic Chips

预组装塑料微流体芯片中人类干细胞衍生神经元的分段化

Use of microfluidic devices to compartmentalize cultured neurons has become a standard method in neuroscience. This protocol shows how to use a ...

Post-differentiation Replating of Human Pluripotent Stem Cell-derived Neurons for High-content Screening of Neuritogenesis and Synapse Maturation

分化后再蛋白的人类多能干细胞衍生神经元，用于高含量的内尿成生和突合性成熟筛查

Neurons differentiated in two-dimensional culture from human pluripotent stem-cell-derived neural progenitor cells (NPCs) represent a powerful model ...

Analyzing Mitochondrial Transport and Morphology in Human Induced Pluripotent Stem Cell-Derived Neurons in Hereditary Spastic Paraplegia

遗传性痉挛性截瘫中人类诱导多能干细胞衍生神经元的线粒体迁移与形态分析

Neurons have intense demands for high energy in order to support their functions. Impaired mitochondrial transport along axons has been observed in human ...

Derivation, Expansion, Cryopreservation and Characterization of Brain Microvascular Endothelial Cells from Human Induced Pluripotent Stem Cells

人类诱导多能干细胞脑微血管内皮细胞的衍生、扩张、冷冻保存和特征化

Brain microvascular endothelial cells (BMECs) can be differentiated from human induced pluripotent stem cells (iPSCs) to develop ex vivo cellular models ...