Name: isolation and culture of hippocampal neurons from prenatal mice
Uploaded: 2012-07-26T12:00:00
Description: Watch this Scientific Journal Video about Isolation and Culture of Hippocampal Neurons from Prenatal Mice at JoVE.com

我们感谢他在帮助准备手稿博士迈克尔·伍滕。这项工作是由NIH 2RO1NS033661（MWW）支持。

1。设置收获前<ol><li>为了产生神经元的收获产前幼仔，安排成年鼠的繁殖19天前神经隔离的日子。 （只C57BL / 6小鼠年龄2-8个月内被用来发展这个协议的目的交配）。在怀孕的女性，心悸或视觉确认，确认成功的交配可以通过检测阴道塞。 </li><li>前一天神经隔离： <ol class="ualpha"><li>对于免疫的应用，在24孔板以3:1的胶原蛋白，鼠尾光涂层的外衣盖玻片：聚-D-赖氨酸的解决方案。 </li><li>用于...

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海马文化已被应用于分子生物学，为20年以上。虽然原则上，神经细胞可以从大脑的任何部分，海马文化已被证明是最流行 ​​，由于在7海马神经细胞的人口相对简单的架构。海马文化通常是由后期的胚胎组织。这个组织是容易分解和包含不少于成熟的脑组织的神经胶质细胞。从胚胎组织的海马神经元的分离，也减少了由于更少的黏附接触3的轴突和树突的剪切破坏。虽然海马的?...

<table border="1"><tbody><tr><td> 试剂名称 </td><td> 供应商 </td><td> 目录编号 </td></tr><tr><td>鼠尾胶原1 </td><td> BD公司</td><td> 354236 </td></tr><tr><td>聚-D-赖氨酸的解决方案</td><td> Chemicon公司</td><td> -003-E的</td></tr><tr><td>汉克斯平衡盐溶液</td><td> Invitrogen公司</td><td> 14175-095 </td></tr><tr><td>胰蛋白酶溶液（1X）0.25％，液体</td><td> Invitrogen公司</td><td> 15050-065 </td></tr><tr><td>的Neurobasal中等液（1X） </td><td> Invitrogen公司</td><td> 21103-049 </td></tr><tr><td> B27的补充（50X）液体</td><td> Invitrogen公司</td><td> 17504-044 </td></tr><tr><td> L-谷氨酰胺200毫米（100X）液体</td><td> Invitrogen公司</td><td> 25030-149 </td></tr><tr><td>青霉素（10,000单位/毫升）/链霉素（10,000微克/毫升） </td><td> Invitrogen公司</td><td> 15140-148 </td></tr><tr><td> HI捐助者马血清</td><td>亚特兰大生物</td><td> S12150H </td></tr></tbody></table>

isolation and culture of hippocampal neurons from prenatal mice

被广泛用于揭示小学文化的大鼠和小鼠海马神经元细胞在神经生物学机制。通过隔离和不断增长的单个神经元，研究人员能够分析贩运细胞，细胞结构和个别蛋白质本地化，使用各种生化技术相关的属性。从这些实验的结果是测试解决学习和记忆功能的神经基础理论的关键。然而，从这些形式的实验结果明确的增长能力，神经细胞与其他脑细胞类型的最低污染的前提。在这个协议中，我们使用特定的神经细胞生长和胚胎海马组织小心剥离，旨在优化健康的神经细胞的生长，同时最大限度地降低污染类型的细胞（即星形胶质细胞）的媒体。胚胎小鼠海马组织能更加困难比DI大小的样品，由于类似的灭鼠组织隔离开来ssection。我们发现从胚胎19天（E19）小鼠幼仔海马的详细解剖技术。一旦海马组织是孤立的，温柔的神经细胞的分离，实现稀浓度的胰蛋白酶和机械设计单独从结缔组织细胞的破坏，同时提供单个细胞的损害最小。包括一个详细说明如何准备用于在中断移液器。免疫荧光协议，以最大限度地提高成功的细胞培养提供最佳的电镀密度。该协议提供了一个快速（约2小时）和有效的技术从小鼠海马组织神经细胞文化。

Watch this Scientific Journal Video about Isolation and Culture of Hippocampal Neurons from Prenatal Mice at JoVE.com

Isolation and Culture of Hippocampal Neurons from Prenatal Mice

我们没有从产前的老鼠大脑海马神经元的神经胶质细胞饲养层的使用高度纯化的文化提供了一个协议。

从产前小鼠海马神经元的分离和培养

Primary cultures of rat and murine hippocampal neurons are widely used to reveal cellular mechanisms in neurobiology. By isolating and growing individual ...

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

Nucleofection and Primary Culture of Embryonic Mouse Hippocampal and Cortical Neurons

Nucleofection和胚胎小鼠海马和皮层神经元的原代培养

Hippocampal and cortical neurons have been used extensively to study central nervous system (CNS) neuronal polarization, axon/dendrite outgrowth, and ...

Calcium Phosphate Transfection of Primary Hippocampal Neurons

原代海马神经元的磷酸钙转染

Calcium phosphate precipitation is a convenient and economical method for transfection of cultured cells. With optimization, it is possible to use this ...

Isolation and Culture of Dissociated Sensory Neurons From Chick Embryos

分离和游离感觉神经元的文化活动从鸡胚

Neurons are multifaceted cells that carry information essential for a variety of functions including sensation, motor movement, learning, and memory. ...

Isolation, Culture and Long-Term Maintenance of Primary Mesencephalic Dopaminergic Neurons From Embryonic Rodent Brains

原发性脑多巴胺能神经元的分离，培养和长期维持胚胎脑鼠类

Degeneration of mesencephalic dopaminergic (mesDA) neurons is the pathological hallmark of Parkinson&#8217;s diseae. Study of the biological processes ...

Low-Density Primary Hippocampal Neuron Culture

低密度原海马神经元文化

The ability to probe the structure and physiology of individual nerve cells in culture is crucial to the study of neurobiology, and allows for flexibility ...

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 ...

Generation of Neurospheres from Mixed Primary Hippocampal and Cortical Neurons Isolated from E14-E16 Sprague Dawley Rat Embryo

从E14-E16斯普拉格·道利大鼠胚胎分离的混合原发希马和皮质神经元生成神经球

Primary neuron culture is an essential technique in the field of neuroscience. To gain deeper mechanistic insights into the brain, it is essential to have ...

Primary Culture of Neurons Isolated from Embryonic Mouse Cerebellum

从胚胎小鼠小脑分离的神经元的主要培养

The use of primary cell cultures has become one of the major tools to study the nervous system in vitro. The ultimate goal of using this simplified model ...

Isolation and Culture of Oculomotor, Trochlear, and Spinal Motor Neurons from Prenatal Islmn:GFP Transgenic Mice

Isolation and Culture of Oculomotor, Trochlear, and Spinal Motor Neurons from Prenatal Islmn:GFP Transgenic Mice

从产前Islmn：GFP转基因小鼠的奥库罗运动、特罗赫利尔和脊柱运动神经元的分离与培养

Oculomotor neurons (CN3s) and trochlear neurons (CN4s) exhibit remarkable resistance to degenerative motor neuron diseases such as amyotrophic lateral ...