我们是从美国国立卫生研究院（奖励R00 NS072192 MKL，HD029178到AS.L.，2 RO1 NS032457到CAW）的支持表示感谢。 MKL是波士顿​​儿童医院的职业发展奖学金/哈佛医学院岸奖学金和研究员Alfred P. Sloan基金会的收件人。 CAW是霍华德·休斯医学研究所的研究员。

1。胚胎分离/准备工作这种技术可以用于小鼠或大鼠。在这个协议中，我们展示了CSF采集技术，大脑皮质的外植体解剖小鼠胚胎脑。我们会就任何重要的差异大鼠与小鼠存在于一般技术。为胚胎的年龄分期系统，E1被归类为天大鼠插头，E0.5被归类为对小鼠的插头的天。 <ol><li>准备一个用Sylgard有机硅弹性体的显微解剖盘。 Sylgard的184有机硅弹性体提供液体成分由两部分组成，第一部分A和B部分，A部分和B部分混合在按重量或体积的比例为10:1。液体组分混合后，倒入一个Petri培养皿中倒入Sylgard的弹性体，以覆盖整个表面的菜。有些气泡可能是本，将在固化过程中消散。将陪替氏培养皿上的盖子，并允许elastom呃治愈。该弹性体可以在室温下固化24小时，或在较高的温度（ 例如 37℃）为更快的固化。一旦液体成分凝固，解剖盘就可以使用了。可反复使用的菜多个实验，只要它被清洁以及下面的过程。 </li><li>的微毛细吸管（针）的制备。制备微毛细吸液管通过施加热和拉使用一个成茂PC-10垂直的微量吸移管拉出器具有以下设置:一个步骤拉加热器＃2设置为58;100克拉砝码。精尖的微管是用细＃55镊子小心折断。针将得到的平均内径为85微米。 </li><li>准备抽吸器组件为CSF愿望。插入微柱塞设置有微毛细吸液管进入毛细管针。或者，将一个塑料的一次性过滤器的端部的吸气管组件，连接吨Ø微毛细吸管。抽吸器组件的另一端的位置上的垫片插入推针贯通。 </li><li>把胚胎从垃圾中分离到微解剖盘准备用Sylgard。 </li><li>移除额外的胚胎外膜和组织，以便清楚地暴露的胚胎。每个组织层第一的子宫壁，然后蜕膜可以解剖用细的虹膜切除剪刀（ 即精细的科学工具＃15000-02）。每个植入部位，首先子宫壁，然后蜕膜可以切开子宫的长轴平行的，并然后被打开切口可进一步用细镊子。蜕膜后，可以删除一个类似的切口，露出胎膜。使不切开或刺破胎膜，应谨慎行使。 </li><li>用无菌Hanks平衡盐溶液（HBSS）清洗，并从周围的胚胎，使用移液管，以及一个kimwipe或过滤器除去过量的流体纸剪成三角形。 </li></ol> 2。心室CSF收集<ol><li>可视化解剖显微镜下鼠标，胚胎应在其一侧铺设，有了一个矢状发育中的胚胎的胚胎。用大鼠胚胎E16或以上的位置，在其背部脊柱的胚胎，沿其最长的平面尺寸，从颅尾方向，在它的后面，如果胚胎是在撒谎。以这种方式收集CSF可以从右侧和左侧侧脑室。 </li><li>稳步微毛细吸管插入到侧脑室，脑脑室或小脑延髓池，试图不接触神经上皮细胞一旦已插入吸管。对于小鼠胚胎E14.5或更旧，被吸出的CSF可以自右心室，然后除去针插入左心室。由于脑室和胚胎的神经管的通畅年龄小于E14.5，整个CSF的体积往往可被吸出从与一个单一的针插入侧脑室。然而，这并非总是如此发育的时间和连接脑室通畅可能略有不同，因此，微毛细吸管也可以被插入到小脑延髓池收集最大CSF体积。 </li><li>一旦微毛细吸管插入侧脑室，中脑脑室或小脑延髓池，仔细，轻轻地开始CSF进入移液管吸移，使用微柱塞创建负压抽吸的CSF，或通过提供一种温柔负压口使得CSF开始轻轻流入微毛细管的吸移管在一个缓慢的和受控的方式创建的。我们建议，观众会与有关机构的政策，这些方法自己的地方官员。 </li><li>继续施加负压，并收集到微毛细吸管的CSF。在小鼠和大鼠胚胎为E16.5/E17或更年轻的，它是可能的轻微上的头部侧的微毛细吸管英寸在较旧的胚胎形成的草皮层的外观，观察心室崩溃，由于大脑的较大尺寸的，它可能不会是可能观察头部的崩溃。 </li><li>停止施加压力，并温和地去除微毛细吸管从侧脑室。 </li><li>轻轻驱逐CSF样品放入Eppendorf管中，已在冰上冷却。 </li><li>从整个一窝动物继续收集CSF和游泳池的样品为同一管。 </li><li>在4℃下以10,000 xg离心10分钟，以除去任何污染的细胞。 </li><li>分析样品的污染神经上皮细胞或红血细胞的任何迹象。污染的迹象，通常表示由流体出现混浊或红/粉红色，或颗粒的存在下，用带血点。离心分离后，将CSF可以通过显微分析的细胞或细胞碎片的任何证据。如果有迹象污染的流体应该被丢弃。作为一个额外的控制，也可被染色的颗粒的含量为细胞。一个干净的CSF样品可用于细胞培养，皮质文化，法，免疫印迹，ELISA，和其他实验。明确CSF应被转移到一个新的无菌Eppendorf管中。现在可以用于分析样品可以与其他样品用液氮冷冻，或管理单元，汇集，并储存在-80℃下冻结和解冻的流体的小样本，可能会导致在小体积减少和蛋白浓度的变化。这可避免通过冷冻干燥的样品。 </li></ol> 3。皮质外植体解剖<ol><li>传输的E14.5胚胎分离出的垃圾到微解剖菜准备用Sylgard。 </li><li>移除胚胎从胚外膜和组织，如在步骤1.5所述。 </li> &lt;李&gt;洗无菌Hanks平衡盐溶液（HBSS），并除去过量的流体，用移液管从周围的胚胎。 </li><li>解剖从胚胎的其余部分（ 图1A）中分离的头部穿过宫颈区域。 </li><li>使用精细的手术刀眼科刀，切下来的中线的头皮。抓住每边用细镊子此切口，去除皮肤。接下来，使用虹膜切除术剪刀做一个切口，运行中线的发展头盖骨的长度。随后，使两个额外的切口，约1/3的距离，从显影颅骨的前部和后部的端部。用细镊子把握"襟翼"的头盖骨，这部分的解剖结果，并拆下显影的头盖骨组织，暴露皮层（ 图1B）。 </li><li>使用手术刀沿中线平分线大脑中期矢状平面中，分离的右侧和左侧的皮质半球（ 图1B，C）。软脑膜和蛛网膜装的皮质，硬脑膜与发展头骨一起被除去。 </li><li>单独准备每个皮质半球。将半球的内侧，以便您可以看到神经节隆起和发展皮层（ 图1C，D）。 </li><li>使用手术刀，使冠状切口，通过大脑皮质神经上皮尾侧嗅球的发展。这的切口应该开始前边界的外侧和内侧神经节隆起，穿过前扣带回区域发展的皮质雏形（ 图1E）。 </li><li>另一个的冠状切口尾部的后边界的横向神经节隆起（ 图1F）。此切口也应该从神经节隆起的侧边界的内侧壁皮质雏形。 </li><li>收回皮质的"啪啪"CREA通过在步骤3.7和3.8的两个切口，使用手术刀或温和的压力从一个流中，用200微升移液器交付的HBSS中，以避免机械损伤的皮质。然后作一横切口，沿边界分离的的横向神经节隆起，从发展皮质（ 图1G，H）。然后，剖析了发展中的海马和大脑皮质的下摆内侧前脑，在新皮层外侧皮质表面符合半球间墙的顶点。 </li><li>作一横切口，沿边界分离的的横向神经节隆起，从皮层（ 图1H）。 </li><li>删除任何额外的Sylgard的板从解剖皮质外植体解剖组织。可以使用温和的移液管移液与新鲜的HBSS缓冲的任何额外的解剖组织。解剖的的皮质是现在有脑膜面朝下（ 图1I）。 </li></ol><ol><li>准备的铂丝的玻璃移液管连接到环路。加热的玻璃移液管与插入的玻璃移液管的端部中的折叠的铂丝。通过加捻的铂丝圈的端部，在循环的大小可以增加或减少，循环可以被成形为适应所需的外植体的大小。 （这可以预先准备和重复使用。） </li><li>的铂丝的端部加热到消毒循环。 </li><li>隔离解剖的皮质外植体并删除额外的的HBSS媒体的移液温和，留少量使用的钢丝环。 </li><li>将1厘米聚碳酸酯膜在4厘米直径的成像碟。 </li><li>使用侧面的铂丝圈，轻轻翻转，使皮层脑膜的一面朝上。 </li><li>关闭夹层皿中，将外植体在循环中间放置和使用的固有的静水压力抬起皮层的外植体电梯外植体在平面上。 </li><li>轻轻地放置在所述聚碳酸酯膜的外植体，并解除铂丝圈离开的外植体，使得仍然心室与膜的表面接触的平坦的平面有上的膜。 </li><li>轻轻抬起膜和膜下吸取50μL的CSF或其它媒体。 </li><li>量的摄像培养皿，将其放置在湿润的副容器，并在5％CO 2在37℃下孵育24小时，以支持持续的细胞增殖和外植体生长。 图2示出了示意性的最终的皮层外植体的培养皿制备。 </li></ol>

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作者宣称，他们没有竞争的金融利益。

心室CSF收集所描述的方法，已经取得了相当纯的胚胎脑脊液样品与在一些细胞分析9的稳定的蛋白质组合物，和一致的活性。有了一个良好的采集技术和产仔数的10 E14.5小鼠，可以预期的CSF收集10-15微升，并从E16鼠一窝，谁也无法预料的CSF收集约50-90微升。此收集技术，从血液和组织中的污染最小化，通过仔细观察，离心分离，和废弃的样品与细胞碎片或血色调的任何证据。我们已经分析了?...

CSF是一种复杂的流体，沐浴神经上皮1-6，并提供必要的压力7和促进生长发育中的大脑8-12线索。研究CSF在大脑发育过程中，我们开发的技术隔离室CSF来自发展中国家的大鼠或小鼠胚胎过程中各阶段的发展6,9。往前的隔离的方法包括用玻璃微针和隔离的CSF使用微喷射器1,2。我们的方法利用玻璃微毛细吸管的尖端一直拉到创建一个超细点，以提高组织的渗透。玻璃微毛细吸管被连接到一个吸气器与温和的压力变化，使心室CSF可以控制收集。为了研究干细胞的的CSF信号的影响，我们解剖大脑皮质外植体，将它们放置在聚碳酸酯膜，并把它们飘浮在适当的铜CSF样品9 lture培养基中。利用这种技术，减少体积的介质来培养的组织，是足够允许脑脊液9的有效利用。

<table border="1"><tbody><tr><td> 的试剂/材料名称 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td> Wiretrop II毛细管针</td><td>德拉蒙德科学</td><td> ＃5-000-2020 </td><td></td></tr><tr><td> SYLGARD </td><td>安士澳粘合剂</td><td> ＃184 SIL ELAST包的</td><td></td></tr><tr><td>吸气管组件</td><td>西格玛</td><td> ＃A5177-5EA </td><td></td></tr><tr><td>一次性过滤器</td><td>文丘里或当地的药店</td><td>不可用</td><td>标准香烟过滤嘴</td></tr><tr><td> Roundstock眼科刀（15度捅一刀） </td><td>世界精密仪器公司</td><td> ＃500250 </td><td></td></tr><tr><td> 35毫米玻璃底培养皿</td><td> MatTek公司</td><td> ＃P35G-1.5-14-C </td><td&gt; </td></tr><tr><td>铂铱丝</td><td> Tritech公司研究</td><td> ＃PT-9010-010-3FT </td><td></td></tr><tr><td>核孔径迹蚀刻膜</td><td> Whatman公司</td><td> ＃09-300-57 </td><td></td></tr><tr><td> Hanks平衡盐溶液</td><td> Fisher Scientific则</td><td> ＃SH30031.FS </td><td></td></tr><tr><td>虹膜切除术剪刀</td><td>精细科学工具</td><td> ＃15000-02 </td><td></td></tr></tbody></table>

isolation of cerebrospinal fluid from rodent embryos for use with dissected cerebral cortical explants

CSF是一种复杂的流体动态变化的蛋白质组的整个开发过程，并在成年后。在胚胎发育过程中，新生的CSF区别于前神经管闭合时的羊水。 CSF卷，然后在随后的日子里，排队的神经上皮干细胞的心室和脉络膜丛产生CSF增加。胚胎CSF接触的顶端，心室发育中的大脑和脊髓神经干细胞的表面。 CSF提供了重要的流体压力发育中的大脑的扩展和分发重要的增长推动因素，神经祖细胞在时间上的具体方式。要调查的CSF的功能，重要的是不从血液或显影端脑组织污染隔离胚胎脑脊液的纯样品。在这里，我们描述了一种技术，以隔离的心室胚胎CSF可用于相对纯的样品质谱，蛋白质电泳，和细胞与初级外植体培养的实验分析，包括范围广泛的。我们演示了如何剖析和文化多孔聚碳酸酯膜的皮层外植体对以成长发展的皮层组织与媒体或CSF的量的减少。使用这种方法，可以进行实验，使用CSF从不同年龄或条件调查对靶细胞的CSF蛋白质组的生物活性。

CSF集合应得到一个明确的，透明的液体。不应该有任何从血液中的污染的证据证明，抽吸，并在Eppendorf管中的流体由一个红色或黄色带点。此外，也应该没有证据表明组织中吸出物和Eppendorf管。当CSF离心，也可以评估的CSF显微镜，以确保有没有污染。如果有污染的迹象时，CSF应该被丢弃。从E14.5鼠标乱抛垃圾，一个可以预见收集10-15微升CSF。 表1描述了从不同胚龄小鼠和大鼠的平均窝?...

Watch this Scientific Journal Video about Isolation of Cerebrospinal Fluid from Rodent Embryos for use with Dissected Cerebral Cortical Explants at JoVE.com

Isolation of Cerebrospinal Fluid from Rodent Embryos for use with Dissected Cerebral Cortical Explants

心室脑脊液（CSF）沐浴在早期的大脑发育在胚胎神经上皮和大脑皮质祖细胞。在这里，我们描述的方法，从不同年龄层的啮齿动物胚胎隔离室CSF，为了研究其生物学功能。此外，我们展示了我们的脑皮质外植体的解剖和文化的技术，可以为外植体生长的培养基或CSF以最小的体积。

脑脊液分离解剖大脑皮质外植体使用的啮齿动物胚胎

The CSF is a complex fluid with a dynamically varying proteome throughout development and in adulthood. During embryonic development, the nascent CSF ...

isolation-cerebrospinal-fluid-from-rodent-embryos-for-use-with

Universidad de Cartagena UDEC

Research

JoVE Journal

Neuroscience

22.9K 次观看.  VA Greater Los Angeles Healthcare System. 心室脑脊液（CSF）沐浴在早期的大脑发育在胚胎神经上皮和大脑皮质祖细胞。在这里，我们描述的方法，从不同年龄层的啮齿动物胚胎隔离室CSF，为了研究其生物学功能。此外，我们展示了我们的脑皮质外植体的解剖和文化的技术，可以为外植体生长的培养基或CSF以最小的体积。

视频: Isolation of Cerebrospinal Fluid from Rodent Embryos for use with Dissected Cerebral Cortical Explants

Mouse Dorsal Forebrain Explant Isolation

This video demonstrates the protocol for isolating and culturing explants of the mouse forebrain from embyonic day 12 mice. Procedures for removal of the uterus, embryos from uterus, and dissection of embryos are given. In addition the methodology for transferring these explants onto specialized membranes on which they are cultured is demonstrated. The development of the forebrain can be studied in vitro using this preparations as well as changes in gene expression.

鼠标背的前脑植隔离

科研

生物学

An Optogenetic Approach for Assessing Formation of Neuronal Connections in a Co-culture System

Here we describe a protocol to generate a co-culture consisting of 2 different neuronal populations. Induced pluripotent stem cells (iPSCs) are reprogrammed from human fibroblasts using episomal vectors. Colonies of iPSCs can be observed 30 days after initiation of fibroblast reprogramming. Pluripotent colonies are manually picked and grown in neural induction medium to permit differentiation into neural progenitor cells (NPCs). iPSCs rapidly convert into neuroepithelial cells within 1 week and retain the capability to self-renew when maintained at a high culture density. Primary mouse NPCs are differentiated into astrocytes by exposure to a serum-containing medium for 7 days and form a monolayer upon which embryonic day 18 (E18) rat cortical neurons (transfected with channelrhodopsin-2 (ChR2)) are added. Human NPCs tagged with the fluorescent protein, tandem dimer Tomato (tdTomato), are then seeded onto the astrocyte/cortical neuron culture the following day and allowed to differentiate for 28 to 35 days. We demonstrate that this system forms synaptic connections between iPSC-derived neurons and cortical neurons, evident from an increase in the frequency of synaptic currents upon photostimulation of the cortical neurons. This co-culture system provides a novel platform for evaluating the ability of iPSC-derived neurons to create synaptic connections with other neuronal populations.

A protocol to generate a co-culture system consisting of neurons derived from induced pluripotent stem cells (iPSCs), primary cortical neurons and astrocytes is described. This co-culture system allows detection of the formation of synaptic contacts and circuits between new, iPSC-derived neurons and pre-existing cortical neurons expressing channelrhodopsin-2.

评定神经连接的形成在共培养体系的光遗传学方法

Here we describe a protocol to generate a co-culture consisting of 2 different neuronal populations. Induced pluripotent stem cells (iPSCs) are ...

发育生物学

Isolation and Culture of Rat Embryonic Neural Cells: A Quick Protocol

We are describing a quick method to dissociate and culture hippocampal or cortical neurons from E15-17 rat embryos. The procedure can be applied successfully to the isolation of mouse and human primary neurons and neural progenitors. Dissociated neurons are maintained in serum-free medium up to several weeks. These cultures can be used for nucleofection, immunocytochemistry, nucleic acids preparation, as well as electrophysiology. Older neuronal cultures can also be transfected with a good efficiency rate by lentiviral transduction and, less efficiently, with calcium phosphate or lipid-based methods such as lipofectamine.

We describe a rapid methodology to isolate and culture hippocampal and cortical neurons from rodent embryos. This protocol allows us to perform experiments in which nearly pure neuronal cultures are required.

大鼠胚胎神经细胞的分离和培养:一个快速议定书"

We are describing a quick method to dissociate and culture hippocampal or cortical neurons from E15-17 rat embryos. The procedure can be applied ...

神经科学

Ex utero Electroporation and Whole Hemisphere Explants: A Simple Experimental Method for Studies of Early Cortical Development

Cortical development involves complex interactions between neurons and non-neuronal elements including precursor cells, blood vessels, meninges and associated extracellular matrix. Because they provide a suitable organotypic environment, cortical slice explants are often used to investigate those interactions that control neuronal differentiation and development. Although beneficial, the slice explant model can suffer from drawbacks including aberrant cellular lamination and migration. Here we report a whole cerebral hemisphere explant system for studies of early cortical development that is easier to prepare than cortical slices and shows consistent organotypic migration and lamination. In this model system, early lamination and migration patterns proceed normally for a period of two days in vitro, including the period of preplate splitting, during which prospective cortical layer six forms. We then developed an ex utero electroporation (EUEP) approach that achieves ~80% success in targeting GFP expression to neurons developing in the dorsal medial cortex.
The whole hemisphere explant model makes early cortical development accessible for electroporation, pharmacological intervention and live imaging approaches. This method avoids the survival surgery required of in utero electroporation (IUEP) approaches while improving both transfection and areal targeting consistency. This method will facilitate experimental studies of neuronal proliferation, migration and differentiation.

Ex utero Electroporation and Whole Hemisphere Explants: A Simple Experimental Method for Studies of Early Cortical Development

This protocol describes an improved explant procedure that involves ex utero electroporation, dissection and culture of entire cerebral hemispheres from the embryonic mouse. The preparation facilitates pharmacological studies and assays of gene function during early cortical development.

前子宫穿孔和整个半球的外植体早期皮质发育的研究:一个简单的实验方法

Cortical development involves complex interactions between neurons and non-neuronal elements including precursor cells, blood vessels, meninges and ...

Hydraulic Extrusion of the Spinal Cord and Isolation of Dorsal Root Ganglia in Rodents

Traditionally, the spinal cord is isolated by laminectomy, i.e. by breaking open the spinal vertebrae one at a time. This is both time consuming and may result in damage to the spinal cord caused by the dissection process. Here, we show how the spinal cord can be extruded using hydraulic pressure. Handling time is significantly reduced to only a few minutes, likely decreasing protein damage. The low risk of damage to the spinal cord tissue improves subsequent immunohistochemical analysis. By performing hydraulic spinal cord extrusion instead of traditional laminectomy, the rodents can further be used for DRG isolation, thereby lowering the number of animals and allowing analysis across tissues from the same rodent. We demonstrate a consistent method to identify and isolate the DRGs according to their localization relative to the costae. It is, however, important to adjust this method to the particular animal used, as the number of spinal cord segments, both thoracic and lumbar, may vary according to animal type and strain. In addition, we illustrate further processing examples of the isolated tissues.

Here, we present a protocol for hydraulic extrusion of the spinal cord as well as identification and isolation of specific dorsal root ganglia (DRGs) in the same rodent. Compared to standard spinal cord isolation methods, this method is significantly faster and reduces the risk of tissue damage.

背根神经节的啮齿动物的脊髓和隔离的液压挤压

Traditionally, the spinal cord is isolated by laminectomy, i.e. by breaking open the spinal vertebrae one at a time. This is both time consuming and may ...

用于神经干细胞和少突胶质细胞祖细胞分离的大鼠"脑挤奶"

The "Brain Milking" Method for the Isolation of Neural Stem Cells and Oligodendrocyte Progenitor Cells from Live Rats

Tissue-specific neural stem cells (NSCs) remain active in the mammalian postnatal brain. They reside in specialized niches, where they generate new neurons and glia. One such niche is the subependymal zone (SEZ; also called the ventricular-subventricular zone), which is located across the lateral walls of the lateral ventricles, adjacent to the ependymal cell layer. Oligodendrocyte progenitor cells (OPCs) are abundantly distributed throughout the central nervous system, constituting a pool of proliferative progenitor cells that can generate oligodendrocytes.
Both NSCs and OPCs exhibit self-renewal potential and quiescence/activation cycles. Due to their location, the isolation and experimental investigation of these cells is performed postmortem. Here, we describe in detail &#34;brain milking&#34;, a method for the isolation of NSCs and OPCs, amongst other cells, from live animals. This is a two-step protocol designed for use in rodents and tested in rats. First, cells are &#34;released&#34; from the tissue via stereotaxic intracerebroventricular (i.c.v.) injection of a &#34;release cocktail&#34;. The main components are neuraminidase, which targets ependymal cells and induces ventricular wall denudation, an integrin-&#946;1-blocking antibody, and fibroblast growth factor-2. At a second &#34;collection&#34; step, liquid biopsies of cerebrospinal fluid are performed from the cisterna magna, in anesthetized rats without the need of an incision.
Results presented here show that isolated cells retain their endogenous profile and that NSCs of the SEZ preserve their quiescence. The denudation of the ependymal layer is restricted to the anatomical level of injection and the protocol (release and collection) is tolerated well&#160; by the animals. This novel approach paves the way for performing longitudinal studies of endogenous neurogenesis and gliogenesis in experimental animals.

作者聚焦:使用新型脑挤奶方案从活体动物中收集神经干细胞和祖细胞 

A method for the isolation of neural stem cells and oligodendrocyte progenitor cells from the brains of live rats is presented here in experimental detail. It allows multiple collections of these cells from the same animals without compromising their well-being.

从活大鼠中分离神经干细胞和少突胶质细胞祖细胞的"脑挤奶"方法

Tissue-specific neural stem cells (NSCs) remain active in the mammalian postnatal brain. They reside in specialized niches, where they generate new ...

A Simple Technique to Isolate and Culture Murine Astrocytes

Source: Wolfes, A. C., et al., <a href="https://appjove.unicartagenaproxy.elogim.com/v/56092">Culturing In Vivo-like Murine Astrocytes Using the Fast, Simple, and Inexpensive AWESAM Protocol.</a> J. Vis. Exp. (2018)
The video demonstrates a simple method to culture murine astrocytes in isolation from other brain cells. During culturing, by using astrocyte-specific media and inhibiting the adherence of non-astrocytic cells, the method selectively cultures astrocytes. These cultured astrocytes exhibit stellate morphology with fine processes.

一种分离和培养小鼠星形胶质细胞的简单技术

All procedures involving sample collection have been performed in accordance with the institute&#39;s IRB guidelines.
1. Rat or Mouse Brain Dissection
...

JoVE Encyclopedia of Experiments

Neuronal Culture Techniques

Primary Neuronal Culture

Isolation and Culturing of Cells from Mouse Embryo Dorsolateral Telencephalons

Source: Landeira, B. S., et al. <a href="https://appjove.unicartagenaproxy.elogim.com/v/56063">Live Imaging of Primary Cerebral Cortex Cells Using a 2D Culture System.</a> J. Vis. Exp. (2017)
This video demonstrates a protocol for isolating and culturing primary cerebral cortex cells from the dorsolateral telencephalon of mouse embryos.

小鼠胚胎背外侧端脑细胞的分离和培养

All procedures involving animal models have been reviewed by the local institutional animal care committee and the JoVE veterinary review board.
1. ...

脑脊液分离解剖大脑皮质外植体使用的啮齿动物胚胎

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此视频中的章节

鼠标背的前脑植隔离

评定神经连接的形成在共培养体系的光遗传学方法

大鼠胚胎神经细胞的分离和培养:一个快速议定书"

前子宫穿孔和整个半球的外植体早期皮质发育的研究:一个简单的实验方法

背根神经节的啮齿动物的脊髓和隔离的液压挤压

从活大鼠中分离神经干细胞和少突胶质细胞祖细胞的"脑挤奶"方法

从活大鼠中分离神经干细胞和少突胶质细胞祖细胞的"脑挤奶"方法

从活大鼠中分离神经干细胞和少突胶质细胞祖细胞的"脑挤奶"方法

一种分离和培养小鼠星形胶质细胞的简单技术

小鼠胚胎背外侧端脑细胞的分离和培养