Name: a functional motor unit in the culture dish: co-culture of spinal cord explants and muscle cells
Uploaded: 2012-04-12T12:00:00
Description: Watch this Scientific Journal Video about A Functional Motor Unit in the Culture Dish: Co-culture of Spinal Cord Explants and Muscle Cells at JoVE.com

支持我们的工作是由瑞士国家科学基金会（民阵），瑞士在系统生物学（SystemsX.ch），美国肌肉萎缩症协会（MDA）含量，协会法国驳LES​​肌病（AFM），美国线粒体疾病基金会倡议（UMDF的），Gebert联阵基金会罕见疾病计划（GRF），瑞士肌肉疾病（SSEM / FSRMM），瑞士人寿“Jubiläumsstiftung献给Volksgesundheit和medizinische Forschung”，罗氏研究基金会和大学的研究学会巴塞尔。

1。小学人体肌肉细胞培养制备<ol><li>建立人体肌肉细胞培养根据对explantation重新explantation的技术4。首先，从非肌肉组织活检的。然后，嵌入在血浆凝块，这使得从外植体成纤维细胞出现1毫米3肌肉植。 </li><li>嵌入植体转入明胶等离子涂层菜，让他们在正常生长介质中成长（的MEM 25％，中型199，10％小牛血清[胎牛血清，1％青霉素/链霉素，10μg/ ml的胰岛素，补充10 ng / ml的重组人?...

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生理体外的工具来研究肌肉细胞的功能，在正常和病理情况下是最高的myologists的利益，因为肌肉细胞培养不重述多种细胞和细胞类型连接的重要性的。增加肌肉细胞纯化的运动神经元是不够的，因为雪旺氏细胞的存在是必需的支配有效率5，它不是拓扑（3D +特约经销）有关的协议描述这里实现功能的运动单位被成功地用于建立神经支配的肌肉细胞， 在体外和在一个完整的NMJ?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td> HBSS中</td><td> GIBCO / Invitrogen公司</td><td> 14170 </td></tr><tr><td>的MEM </td><td> GIBCO / Invitrogen公司</td><td> 31095 </td></tr><tr><td>中等199 </td><td> GIBCO / Invitrogen公司</td><td> 31153 </td></tr><tr><td>胎牛血清</td><td>胚胎克隆Perbio </td><td> SH30066.03 </td></tr><tr><td>胰岛素</td><td>西格玛</td><td> I9278 </td></tr><tr><td>人类表皮生长因子</td><td>西格玛</td><td> E9644 </td></tr><tr><td>人类FGF </td><td>硅GMA </td><td> F0291 </td></tr><tr><td>青霉素/链霉素溶液</td><td> Gibco公司</td><td> 15140 </td></tr></tbody></table>

a functional motor unit in the culture dish: co-culture of spinal cord explants and muscle cells

人类的主要肌肉细胞单层培养aneurally很少自发的，因为在一个神经组件的情况下，细胞分化是有限的和运动神经元的刺激，失踪1人的合同。这些限制妨碍在体外培养的肌肉细胞中的许多神经肌肉疾病的研究。重要的是，单层，培养肌细胞的实验限制是可以克服的功能神经支配的肌纤维与脊髓外植体，在联合培养。 在这里，我们实现了高效，正确支配人类的主要肌肉细胞所需的不同步骤，根据Askanas 2开发的方法，导致完成分化和纤维收缩。这样做，肌肉细胞共同培养大鼠胚胎脊髓植13.5海关，背根节还连着脊髓切片。几天后，肌肉FIBERS开始合同，并最终成为通过支配跨横纹肌从外植体，连接到肌肉细胞的脊髓投射功能的突起。这种结构可以维持很多个月，只需定期交流培养基。 这宝贵的工具的应用有很多，因为它代表了一种对人体肌肉的发展和支配的多学科分析的功能模型。事实上，一个完整的从头神经肌肉交界处安装发生在培养皿中，让每一步，从根本上和生理范围内，一个容易的多参数测量。 只是举出几个例子，基因和/或蛋白质的研究，可以直接进行联合培养。此外，前和突触后的效果可以明确，并分别在神经肌肉接头处评估，从不同的物种，因为这两个组件，老鼠和人，分别为。共培养神经肌肉也可以从肌肉或神经肌肉疾病患者分离出人体肌肉细胞进行，因而可以作为候选药物的筛选工具。最后，没有特殊的设备，但需要定期BSL2设施重现功能的运动单位在培养皿。因此，这种方法是有价值的肌肉以及肌肉神经肌肉功能的生理和机理研究研究社区，在正常和疾病的情况下。

Watch this Scientific Journal Video about A Functional Motor Unit in the Culture Dish: Co-culture of Spinal Cord Explants and Muscle Cells at JoVE.com

A Functional Motor Unit in the Culture Dish: Co-culture of Spinal Cord Explants and Muscle Cells

体外培养肌细胞是不够的模型来概括神经支配的肌肉<em&gt;在体内</em&gt;。可以复制功能的运动单位<em&gt;在体外</em&gt;分化人类的主要肌肉细胞用鼠胚胎脊髓植支配。本文介绍了如何建立脊髓植和肌肉细胞共培养。

一个在培养皿中的功能电机组：脊髓植和肌肉细胞共培养

Human primary muscle cells cultured  aneurally in monolayer rarely contract spontaneously because, in the absence of  a nerve component, cell ...

Research

JoVE Journal

Neuroscience

Dissection and Culture of Commissural Neurons from Embryonic Spinal Cord

从胚胎脊髓​​连合神经的解剖和文化

Commissural neurons have been widely used to investigate the mechanisms underlying axon guidance during embryonic spinal cord development. The cell bodies ...

科研

神经科学

Dissection and Culture of Chick Statoacoustic Ganglion and Spinal Cord Explants in Collagen Gels for Neurite Outgrowth Assays

小鸡Statoacoustic神经节和脊髓植在胶原凝胶夹层和文化突起生长实验

The sensory organs of the chicken inner ear are innervated by the peripheral processes of statoacoustic ganglion (SAG) neurons. Sensory organ innervation ...

Chicken Embryo Spinal Cord Slice Culture Protocol

鸡胚胎脊髓片培养协议

Slice cultures can facilitate the manipulation of  embryo development both pharmacologically and through gene manipulations. In  this reduced system, ...

Analysis of Developing Tooth Germ Innervation Using Microfluidic Co-culture Devices

微流控采用共培养开发设备牙胚支配分析

Innervation plays a key role in the development, homeostasis and regeneration of organs and tissues. However, the mechanisms underlying these phenomena ...

Zebrafish In Situ Spinal Cord Preparation for Electrophysiological Recordings from Spinal Sensory and Motor Neurons

Zebrafish In Situ Spinal Cord Preparation for Electrophysiological Recordings from Spinal Sensory and Motor Neurons

斑马鱼<em&gt;原位</em&gt;脊髓制备用于脊髓感觉和运动神经元的电生理记录

Zebrafish, first introduced as a developmental model, have gained popularity in many other fields. The ease of rearing large numbers of rapidly developing ...

Spinal Cord Neurons Isolation and Culture from Neonatal Mice

脊髓神经元从新生小鼠中分离和培养

We present a protocol for the isolation and culture of spinal cord neurons. The neurons are obtained from neonatal C57BL/6 mice and are isolated on ...

Laminectomy and Spinal Cord Window Implantation in the Mouse

拉米内切手术和脊髓窗口植入小鼠

This protocol describes a method for spinal cord laminectomy and glass window implantation for in vivo imaging of the mouse spinal cord. An integrated ...

Identifying Cell Surface Markers of Primary Neural Stem and Progenitor Cells by Metabolic Labeling of Sialoglycan

通过Sialoglycan的代谢标记识别主要神经干细胞和祖细胞的细胞表面标记

Neural stem and progenitor cells (NSPCs) are the cellular basis for the complex structures and functions of the brain. They are located in specialized ...

Direct Injection of a Lentiviral Vector Highlights Multiple Motor Pathways in the Rat Spinal Cord

直接注射慢病毒载体突出大鼠脊髓中的多种运动途径

Introducing proteins of interest into cells in the nervous system is challenging due to innate biological barriers that limit access to most molecules. ...

Conversion of Human Induced Pluripotent Stem Cells (iPSCs) into Functional Spinal and Cranial Motor Neurons Using PiggyBac Vectors

Conversion of Human Induced Pluripotent Stem Cells iPSCs into Functional Spinal and Cranial Motor Neurons Using PiggyBac Vectors

利用 PiggyBac 载体将人诱导的多能干细胞 (Ipsc) 转化为功能性脊柱和颅内运动神经元

We describe here a method to obtain functional spinal and cranial motor neurons from human induced pluripotent stem cells (iPSCs). Direct conversion into ...