Name: culture of neurospheres derived from the neurogenic niches in adult prairie voles
Uploaded: 2020-06-10T14:45:00
Description: Watch this Scientific Journal Video about Culture of Neurospheres Derived from the Neurogenic Niches in Adult Prairie Voles at JoVE.com

这项研究得到了国家委员会252756和253631赠款的支持;联南特派团-DGAPA-PAPIIT IN202818和203518;INPER 2018-1-163，NIH P51OD11132。我们感谢迪西·加斯卡、卡洛斯·洛扎诺、马丁·加西亚、亚历杭德拉·卡斯蒂利亚、尼迪亚·埃尔南德斯、杰西卡·诺里斯和苏珊娜·卡斯特罗的出色技术援助。

这项研究得到了墨西哥墨西哥国家国家大学神经生物学研究所研究伦理委员会和全国生物学研究所（2018-1-163）的批准。这些动物的繁殖、护理和人道终点是根据墨西哥卫生秘密协会"萨卢德总健康法"（卫生研究一般健康法）根据墨西哥官方标准（NOM-062-Z00-1999）建立的。
1. 解决方案和库存准备
<ol><li>准备N2培养基与485 mL的Dulbecco的修改鹰中-F12（DMEM-F12），5mL的N2补充剂（100x）?...

<ol>
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G., 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=Pup+exposure+elicits+hippocampal+cell+proliferation+in+the+prairie+vole.">Pup exposure elicits hippocampal cell proliferation in the prairie vole.</a> Behavioral Brain Research. 187 (1), 9-16 (2008).</li><li>Wojtowicz, J. M., Kee, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=BrdU+assay+for+neurogenesis+in+rodents.">BrdU assay for neurogenesis in rodents.</a> Nature Protocols. 1 (3), 1399-1405 (2006).</li><li>Eack, S. M., 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=Commonalities+in+social+and+non-social+cognitive+impairments+in+adults+with+autism+spectrum+disorder+and+schizophrenia.">Commonalities in social and non-social cognitive impairments in adults with autism spectrum disorder and schizophrenia.</a> Schizophrenia Research. 148 (1-3), 24-28 (2013).</li><li>Pinkham, A. E., 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=Comprehensive+comparison+of+social+cognitive+performance+in+autism+spectrum+disorder+and+schizophrenia.">Comprehensive comparison of social cognitive performance in autism spectrum disorder and schizophrenia.</a> Psychological Medicine. , 1-9 (2019).</li><li>Yirmiya, N., 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=Association+between+the+arginine+vasopressin+1a+receptor+(AVPR1a)+gene+and+autism+in+a+family-based+study:+mediation+by+socialization+skills.">Association between the arginine vasopressin 1a receptor (AVPR1a) gene and autism in a family-based study: mediation by socialization skills.</a> Molecular Psychiatry. 11 (5), 488-494 (2006).</li><li>Montag, C., 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=Oxytocin+and+oxytocin+receptor+gene+polymorphisms+and+risk+for+schizophrenia:+a+case-control+study.">Oxytocin and oxytocin receptor gene polymorphisms and risk for schizophrenia: a case-control study.</a> The World Journal of Biological Psychiatry. 14 (7), 500-508 (2013).</li><li>Harony, H., Wagner, S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+contribution+of+oxytocin+and+vasopressin+to+mammalian+social+behavior:+potential+role+in+autism+spectrum+disorder.">The contribution of oxytocin and vasopressin to mammalian social behavior: potential role in autism spectrum disorder.</a> Neurosignals. 18 (2), 82-97 (2010).</li><li>Bachner-Melman, R., Ebstein, R. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+role+of+oxytocin+and+vasopressin+in+emotional+and+social+behaviors.">The role of oxytocin and vasopressin in emotional and social behaviors.</a> Handbook of Clinical Neurology. 124, 53-68 (2014).</li><li>Wegiel, J., 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=The+neuropathology+of+autism:+defects+of+neurogenesis+and+neuronal+migration,+and+dysplastic+changes.">The neuropathology of autism: defects of neurogenesis and neuronal migration, and dysplastic changes.</a> Acta Neuropathologica. 119 (6), 755-770 (2010).</li><li>Kaushik, G., Zarbalis, K. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Prenatal+Neurogenesis+in+Autism+Spectrum+Disorders.">Prenatal Neurogenesis in Autism Spectrum Disorders.</a> Frontiers in Chemistry. 4, 12 (2016).</li><li>Sheu, J. 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=A+Critical+Period+for+the+Development+of+Schizophrenia-Like+Pathology+by+Aberrant+Postnatal+Neurogenesis.">A Critical Period for the Development of Schizophrenia-Like Pathology by Aberrant Postnatal Neurogenesis.</a> Frontiers in Neuroscience. 13, 635 (2019).</li><li>Donaldson, Z. R., Young, L. 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获得神经干细胞培养的阶段是酶溶液的消化期，不应超过30分钟，因为它可能会降低细胞的生存能力。神经球应在初始培养后8-10天出现;如果第12天没有出现，就放弃培养，重复实验，缩短消化期。另一个问题就是覆盖脑组织的血管。在解剖过程中，它们应该被完全去除，因为红细胞的过量会干扰神经球的形成。
该协议允许扩展浮动神经球，直到通过2和改变粘附条件，以评估?...

草原卷（Microtus ochrogaster）是克里西蒂达家族的一员，是一种小型哺乳动物，其生命策略发展为社会一夫一妻制和高度善于交际的物种。雄性与雌性在交配或长期同居后建立持久的对结，其特点是共享巢，保卫自己的领地，并表现出双亲照顾他们的后代1，2，3，4。因此，草原卷是理解社会性行为和社会认知障碍的神经生物学基础的宝贵模型。
成人神经生成是导致行为变化的神经可塑性最重要的过程之一。例如，我们的研究小组在雄细胞中报告说，与交配的社会同居增加了在海马体凹陷陀螺（DG）的辅助区（VZ）和亚角区细胞增殖，这表明成人神经生成可以在草原卷交配诱导的配对结合（未公布的数据）中发挥作用。另一方面，虽然产生和整合新神经元的大脑区域是众所周知的，但由于整个大脑模型6的技术缺点，这些过程中涉及的分子和细胞机制仍然不确定。例如，控制基因表达和其他细胞活动的信号通路具有相对较短的激活期（磷蛋白群的检测）7。另一种模型是分离和培养成人神经干细胞或祖细胞，以阐明参与成人神经生成分子成分。
维持成年哺乳动物（小鼠）大脑中的神经前体的第一个方法是神经球的测定，神经球是在非粘附条件下生长的细胞聚集体，保留了其产生神经元的多能潜力，以及星细胞8、9、10。在它们的发展过程中，有一个选择过程，只有前体会响应米粒生长因子（EGF）和纤维细胞生长因子2（FGF2）等表皮生长因子（FGF2），以增殖和产生神经球8，9，10。
据我们所知，文献中没有关于从草原卷获得成人神经祖细胞的协议。在这里，我们建立了培养条件，通过神经球形成分析将神经元祖细胞从神经原位及其体外维持中分离。因此，实验可以设计为确定神经干细胞和祖细胞的增殖、迁移、分化和存活的分子和细胞机制，这些机制在草原卷中仍然不为人知。此外，阐明从VZ和DG衍生的细胞的特性的体外差异，可以提供神经基因利基在神经可塑性中的作用的信息，与社会性行为和认知行为的变化，以及社会互动的不足（自闭症谱系障碍和精神分裂症），这也可能依赖于性别。

<table>
	<tbody>
		<tr>
			<td>Antibodies</td>
			<td></td>
			<td></td>
			<td>Antibody ID</td>
		</tr>
		<tr>
			<td>Anti-Nestin</td>
			<td>GeneTex</td>
			<td>GTX30671</td>
			<td>RRID:AB_625325</td>
		</tr>
		<tr>
			<td>Anti-Doublecortin</td>
			<td>MERCK</td>
			<td>AB2253</td>
			<td>RRID:AB_1586992</td>
		</tr>
		<tr>
			<td>Anti-Ki67</td>
			<td>Abcam</td>
			<td>ab66155</td>
			<td>RRID:AB_1140752</td>
		</tr>
		<tr>
			<td>Anti-MAP2</td>
			<td>GeneTex</td>
			<td>GTX50810</td>
			<td>RRID:AB_11170769</td>
		</tr>
		<tr>
			<td>Anti-GFAP</td>
			<td>SIGMA</td>
			<td>G3893</td>
			<td>RRID:AB_477010</td>
		</tr>
		<tr>
			<td>Goat Anti-Mouse Alexa Fluor 488</td>
			<td>Thermo Fisher Scientific</td>
			<td>A-11029</td>
			<td>RRID:AB_2534088</td>
		</tr>
		<tr>
			<td>Goat Anti-Rabbit Alexa Fluor 568</td>
			<td>Thermo Fisher Scientific</td>
			<td>A-11036</td>
			<td>RRID:AB_10563566</td>
		</tr>
		<tr>
			<td>Goat Anti-Guinea Pig Alexa Fluor 488</td>
			<td>Thermo Fisher Scientific</td>
			<td>A-11073</td>
			<td>RRID:AB_2534117</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Culture reagents</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Antibiotic-Antimycotic</td>
			<td>Thermo Fisher Scientific/Gibco</td>
			<td>15240062</td>
			<td>100X</td>
		</tr>
		<tr>
			<td>B-27 supplement</td>
			<td>Thermo Fisher Scientific/Gibco</td>
			<td>17504044</td>
			<td>50X</td>
		</tr>
		<tr>
			<td>Collagenase, Type IV</td>
			<td>Thermo Fisher Scientific/Gibco</td>
			<td>17104019</td>
			<td>Powder</td>
		</tr>
		<tr>
			<td>Dispase</td>
			<td>Thermo Fisher Scientific/Gibco</td>
			<td>17105041</td>
			<td>Powder</td>
		</tr>
		<tr>
			<td>DMEM/F12, HEPES</td>
			<td>Thermo Fisher Scientific/Gibco</td>
			<td>11330032</td>
			<td></td>
		</tr>
		<tr>
			<td>Glucose</td>
			<td>any brand</td>
			<td></td>
			<td>Powder, Cell Culture Grade</td>
		</tr>
		<tr>
			<td>GlutaMAX</td>
			<td>Thermo Fisher Scientific/Gibco</td>
			<td>35050061</td>
			<td>100X</td>
		</tr>
		<tr>
			<td>HEPES</td>
			<td>any brand</td>
			<td></td>
			<td>Powder, Cell Culture Grade</td>
		</tr>
		<tr>
			<td>Mouse Laminin</td>
			<td>Corning</td>
			<td>354232</td>
			<td>1 mg/mL</td>
		</tr>
		<tr>
			<td>N-2 supplement</td>
			<td>Thermo Fisher Scientific/Gibco</td>
			<td>17502048</td>
			<td>100X</td>
		</tr>
		<tr>
			<td>NAHCO3</td>
			<td>any brand</td>
			<td></td>
			<td>Powder, Suitable for Cell Culture</td>
		</tr>
		<tr>
			<td>Neurobasal</td>
			<td>Thermo Fisher Scientific/Gibco</td>
			<td>21103049</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate-Buffered Saline (PBS)</td>
			<td>Thermo Fisher Scientific/Gibco</td>
			<td>10010023</td>
			<td>1X</td>
		</tr>
		<tr>
			<td>Poly-L-ornithine hydrobromide</td>
			<td>Sigma-Aldrich</td>
			<td>P3655</td>
			<td>Powder</td>
		</tr>
		<tr>
			<td>Recombinant Human EGF</td>
			<td>Peprotech</td>
			<td>AF-100-15</td>
			<td></td>
		</tr>
		<tr>
			<td>Recombinant Human FGF-basic</td>
			<td>Peprotech</td>
			<td>AF-100-18B</td>
			<td></td>
		</tr>
		<tr>
			<td>StemPro Accutase Cell Dissociation Reagent</td>
			<td>Thermo Fisher Scientific/Gibco</td>
			<td>A1110501</td>
			<td>100 mL</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Disposable material</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>24-well Clear Flat Bottom Ultra-Low Attachment Multiple Well Plates</td>
			<td>Corning/Costar</td>
			<td>3473</td>
			<td></td>
		</tr>
		<tr>
			<td>24-well Clear TC-treated Multiple Well Plates</td>
			<td>Corning/Costar</td>
			<td>3526</td>
			<td></td>
		</tr>
		<tr>
			<td>40 &#181;m Cell Strainer</td>
			<td>Corning/Falcon</td>
			<td>352340</td>
			<td>Blue</td>
		</tr>
		<tr>
			<td>Bottle Top Vacuum Filter, 0.22 &#181;m pore</td>
			<td>Corning</td>
			<td>431118</td>
			<td>PES membrane, 45 mm diameter neck</td>
		</tr>
		<tr>
			<td>Non-Pyrogenic Sterile Centrifuge Tube</td>
			<td>any brand</td>
			<td></td>
			<td>with conical bottom</td>
		</tr>
		<tr>
			<td>Non-Pyrogenic sterile tips of 1,000 &#181;l, 200 &#181;l and 10 &#181;l.</td>
			<td>any brand</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile cotton gauzes</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile microcentrifuge tubes of 1.5 mL</td>
			<td>any brand</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile serological pipettes of 5, 10 and 25 mL</td>
			<td>any brand</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Sterile surgical gloves</td>
			<td>any brand</td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Syringe Filters, 0.22 &#181;m pore</td>
			<td>Merk Millipore</td>
			<td>SLGPR33RB</td>
			<td>Polyethersulfone (PES) membrane, 33 mm diameter</td>
		</tr>
		<tr>
			<td></td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Equipment and surgical instruments</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Biological safety cabinet</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dissecting Scissors</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Dumont Forceps</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Motorized Pipet Filler/Dispenser</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Micropipettes</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Petri Dishes</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Scalpel Blades</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
		<tr>
			<td>Stainless-steel Spatula</td>
			<td></td>
			<td></td>
			<td></td>
		</tr>
	</tbody>
</table>

culture of neurospheres derived from the neurogenic niches in adult prairie voles

神经球是组成神经干细胞和祖细胞的原发细胞聚集体。这些 3D 结构是确定神经干细胞的分化和增殖潜力以及生成细胞系的极佳工具。此外，神经球可以创建一个利基（体外），允许动态变化的环境建模，如不同的生长因子，激素，神经递质，等等。 微图色（ 草原卷）是理解社会性行为和社会认知的神经生物学基础的独特模型。然而，这些行为所涉及的细胞机制并不为人所知。该协议旨在从成人草原卷的神经原位获得神经祖细胞，这些细胞在非粘附条件下培养，以产生神经球。神经球的大小和数量取决于区域（辅助区或凹陷陀螺）和草原卷的性别。这种方法是研究体外神经基因利基的性别依赖差异和与社会行为相关的神经可塑性变化（如配对结合和双亲护理）的显著工具。此外，可以检查导致社会互动缺陷的认知条件（自闭症谱系障碍和精神分裂症）。

神经球是由从雌雄成年草原卷的VZ和DG分离出的神经干细胞形成的。开始培养后大约8-10天，细胞应该已经形成神经球。请注意，该板可能包含主培养物中的碎屑（图 3A）。然而，在第1段，培养者应该只包括神经球（图3B）。
与雌性VZ和雌性VZ相比，从女性VZ获得的神经球数量更高（图4A）。这些数据表明，获得?...

Watch this Scientific Journal Video about Culture of Neurospheres Derived from the Neurogenic Niches in Adult Prairie Voles at JoVE.com

Culture of Neurospheres Derived from the Neurogenic Niches in Adult Prairie Voles

我们建立了培养神经祖细胞的条件，从辅助区和凹陷陀螺的成年大脑的草原卷，作为补充的体外研究，以分析神经基因利基之间的性别依赖差异，这可能是功能塑料变化的一部分相关的社会行为。

从成人草原卷的神经性尼奇衍生的神经球培养

Neurospheres are primary cell aggregates that comprise neural stem cells and progenitor cells. These 3D structures are an excellent tool to determine the ...

这种方法可以帮助研究神经原性利基的性别依赖差异，并了解它们的神经可塑性，这些变化突出了草原中的社会互动。神经球的测定是确定神经干细胞和祖细胞增殖和分化潜力的极好工具。首先，在被冰包围的表面上放置一个培养皿。将大脑沉积在盘子上，加入20毫升的冷洗溶液。在日冕平面上，用手术刀将大脑分成两个组织块，在前后轴的脑力水平上执行切割。从骨块中提取辅助区或VZ组织，从骨块中提取凹陷陀螺（或DG）组织。要解剖VZ，请用杜蒙钳子握住半球之一，并在组织下插入第二钳子的细尖，这些钳子与卡达特-普塔门一起行。沿背心-心轴打开钳子以分离组织，将 VZ 收集到具有两毫升冷洗溶液的离心管中。不要将两种动物的组织集中。在另一半球重复微切除，将管与组织储存在冰上。要解剖DG，使用手术刀将冠状切入冠状块，获得两片，其中海马形成被观察。使用 Dumont 钳子握住其中一个切片，并在 DG 和 CA1 之间用另一个钳子进行水平切割。然后，在 DG 和 CA3 之间进行垂直切口，以分离 DG。在第一个切片的另一个半球中重复解剖，然后在第二个切片中的两个半球重复解剖。在离心管中收集每卷的四个 DG 片。将离心管放在生物安全柜内，等待大约 10 分钟，让组织碎片被重力沉淀。然后，取出洗涤液，并添加一毫升温暖的酶溶液到每个管。在37度下孵育管子10分钟。要分解组织片段，用一毫升的尖端上下移液，但移液器不超过30次。在37摄氏度下重复孵育，然后再次移液组织。在每个管中加入9毫升N-2介质以稀释酶处理，并将管以200倍g离心4分钟。丢弃上经剂，用10毫升N-2介质清洗细胞，然后重复离心。从每个管中去除上一提液，并分别将VZ和DG的细胞颗粒用两毫升和一毫升的B-27介质重新暂停。用40微米细胞滤株过滤每个细胞悬浮液，以去除任何非消化组织。培养超低附着24孔板中的过滤细胞，使用两口用于VZ的孔和一口用于DG的孔。每毫升成纤维细胞生长因子20毫微克，每毫升表皮生长因子20毫微克，然后在37摄氏度、5%二氧化碳和高湿度下孵育48小时。孵育后，去除一半的培养基，代之以新鲜的B-27培养基，辅以双浓度的生长因子。每三天重复一次此过程。在不需要改变培养媒介的日子里，将生长因子添加到最终浓度为 1 倍。神经球是由从辅助区分离出的神经干细胞和雌雄成年草原卷的凹陷陀螺形成。虽然在初级培养物中存在碎片，但在第一次通过后，只有神经球存在。从女性辅助区获得的神经球数量比从男性补助区获得的神经球数量多，或雌性或男性的凹陷陀螺，这表明获得的神经球数量取决于增殖区和卷性。神经球的直径在8天、第11天和14天测量。在两个神经元区域，男性和女性的卷逐渐增加，但与来自女性大脑的神经球相比，来自男性大脑的神经球较小。粘附神经球在第六天有生长因子时出现特征，或第15天无生长因子。在第六天，在未分化的条件下，神经球衍生细胞表达内丁，神经祖细胞的标记。也可以识别双皮质蛋白阳性细胞和增殖标记Ki-67，这表明存在神经元前体或不成熟的神经元。然而，Ki-67与DCX缺乏结肠化表明存在后线粒体神经细胞。在第15天，在分化条件下，发现成熟的神经元和细胞与胶质表型，证明分离细胞的分化潜力。在尝试此协议时，请记住，能够识别和有以前在微切除方面的实践是将细胞从神经原性利基中分离出来的根本。按照这个协议，实验可以设计成与神经干细胞和祖细胞的增殖、分化和存活有关的机制，这些机制在草原卷中仍然不为人知。

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

Organotypic Culture of Full-thickness Adult Porcine Retina

器官型培养的全厚成年猪视网膜

There is a recognized demand for in vitro models that can replace or reduce animal experiments. Porcine retina has a similar neuronal structure to human ...

Progenitor-derived Oligodendrocyte Culture System from Human Fetal Brain

来自祖，人脑少突胶质细胞培养系统

Differentiation of human neural progenitors into neuronal and glial cell types offers a model to study and compare molecular regulation of neural cell ...

An In-vitro Preparation of Isolated Enteric Neurons and Glia from the Myenteric Plexus of the Adult Mouse

An In-vitro Preparation of Isolated Enteric Neurons and Glia from the Myenteric Plexus of the Adult Mouse

一个<em在体外</em从成年小鼠的肌间神经丛制备隔离肠道神经元和神经胶质

The enteric nervous system is a vast network of neurons and glia running the length of the gastrointestinal tract that functionally controls ...

One Mouse, Two Cultures: Isolation and Culture of Adult Neural Stem Cells from the Two Neurogenic Zones of Individual Mice

一个鼠标，两种文化：隔离和成人神经的培养干细胞从单个小鼠的两个神经源性区

The neurosphere assay and the adherent monolayer culture system are valuable tools to determine the potential (proliferation or differentiation) of adult ...

Lineage-reprogramming of Pericyte-derived Cells of the Adult Human Brain into Induced Neurons

谱系重编程的成人大脑的周细胞源性细胞诱导成神经细胞

Direct lineage-reprogramming of non-neuronal cells into induced neurons (iNs) may provide insights into the molecular mechanisms underlying neurogenesis ...

Isolation and Culture of Adult Neural Stem Cells from the Mouse Subcallosal Zone

分离和成人神经干细胞的培养从鼠标Subcallosal区

Adult neural stem cells (aNSCs) can be used for the regeneration of damaged brain tissue. NSCs have the potential for differentiation and proliferation ...

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

A Simple One-step Dissection Protocol for Whole-mount Preparation of Adult Drosophila Brains

一个简单的一步解剖协议成人全安装准备<em&gt;果蝇</em&gt;脑

There is an increasing interest in using Drosophila to model human brain degenerative diseases, map neuronal circuitries in adult brains, and study the ...

Simple Generation of a High Yield Culture of Induced Neurons from Human Adult Skin Fibroblasts

人类成人皮肤成纤维细胞诱导神经元高产培养的简单生成

Induced neurons (iNs), the product of somatic cells directly converted to neurons, are a way to obtain patient-derived neurons from tissue that is easily ...

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 ...