Name: the neuroblast assay: an assay for the generation and enrichment of neuronal progenitor cells from differentiating neural stem cell progeny using flow cytometry
Uploaded: 2012-04-22T14:00:00
Description: Watch this Scientific Journal Video about The Neuroblast Assay: An Assay for the Generation and Enrichment of Neuronal Progenitor Cells from Differentiating Neural Stem Cell Progeny Using Flow Cytomet

这项工作是从斯特里特基金会的资金支持。

1。细胞培养之前基本设置<ol><li>国科会NeuroCult基础培养基和NeuroCult国科会扩散补充，以9:1的比例混合，分别作出完整的国科会介质。 </li><li> 37℃水浴用于热身介质。 </li><li>适量热灭活胎牛血清（FCS）的解冻。 </li><li>适量股票生长因子解决方案，包括表皮生长因子（表皮生长因子在10微克/毫升），碱性成纤维细胞生长因子（b-FGF的10微克/毫升）和0.2％肝素溶液磷酸缓冲液（PBS）解冻。 </li><li>组?...

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	<li>Azari, H., Rahman, M., Sharififar, S., Reynolds, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+Expansion+of+the+Adult+Mouse+Neural+Stem+Cells+Using+the+Neurosphere+Assay.">Isolation and Expansion of the Adult Mouse Neural Stem Cells Using the Neurosphere Assay.</a> J. Vis. Exp. (45), e2393 (2010).</li><li>Azari, H., Sharififar, S., Rahman, M., Ansari, S., Reynolds, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Establishing+Embryonic+Mouse+Neural+Stem+Cell+Culture+Using+the+Neurosphere+Assay.">Establishing Embryonic Mouse Neural Stem Cell Culture Using the Neurosphere Assay.</a> J. Vis. Exp. (47), e2457 (2011).</li><li>Siebzehnrubl, F. A., Vedam-Mai, V., Azari, H., Reynolds, B. A., Deleyrolle, L. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+Characterization+of+Adult+Neural+Stem+Cells.">Isolation and Characterization of Adult Neural Stem Cells.</a> Methods. Mol. Biol. 750, 61-77 (2011).</li><li>Scheffler, B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Phenotypic+and+functional+characterization+of+adult+brain+neuropoiesis.">Phenotypic and functional characterization of adult brain neuropoiesis.</a> Proceedings of the National Academy of Sciences of the United States of America. 102, 9353-9358 (2005).</li><li>Azari, H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Purification+of+immature+neuronal+cells+from+neural+stem+cell+progeny.">Purification of immature neuronal cells from neural stem cell progeny.</a> PLoS One. 6, e20941-e20941 (2011).</li><li>Amariglio, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Donor-Derived+Brain+Tumor+Following+Neural+Stem+Cell+Transplantation+in+an+Ataxia+Telangiectasia+Patient.">Donor-Derived Brain Tumor Following Neural Stem Cell Transplantation in an Ataxia Telangiectasia Patient.</a> Plos Medicine. 6, 221-231 (2009).</li></ol>

隔离定义的神经细胞群（即神经元，星形胶质细胞和少突胶质细胞）的能力可能会解决一些与临床应用神经干细胞（NS​​Cs）的相关障碍。首先，它使我们的供体细胞的人口开始充分体现。第二，它可以让我们使用一个特定的细胞类型或不同类型的细胞组合，与特定的比例，根据疾病的性质或阶段。最后，使用高浓缩铀的预分化的神经细胞的祖细胞，可以显着降低可能不受控制地增殖和肿瘤的形...

<table border="1"><tr><td> 试剂名称 </td><td> 类型 </td><td> 公司 </td><td> 目录编号 </td><td> 评论 </td></tr><tr><td> NeuroCult国科会的基础培养基</td><td>中等</td><td>干细胞技术</td><td> 05700 </td><td></td></tr><tr><td> NeuroCult国科会扩散补充</td><td>中等补充</td><td>干细胞技术</td><td> 05701 </td><td></td></tr><tr><td> ％0.05胰蛋白酶EDTA </td><td>试剂</td><td> Gibco公司</td><td> 25300-062 </td><td></td></tr><tr><td>大豆胰蛋白酶抑制剂</td><td>试剂</td><td>西格玛</td><td> T6522 </td><td></td></tr><tr><td>笔/链球菌</td><td>试剂</td><td> Gibco公司</td><td> 15140-122 </td><td></td></tr><tr><td> *的MEM </td><td>试剂</td><td> Gibco公司</td><td> 41500-018 </td><td> HEM的组成部分</td></tr><tr><td> *肝素钠</td><td>试剂</td><td>西格玛</td><td> H4034 </td><td> HEM的组成部分</td></tr><tr><td> *蒸馏水</td><td>试剂</td><td> Gibco公司</td><td> 15230-147 </td><td></td></tr><tr><td>细胞过滤器</td><td>筛</td><td>屋宇署猎鹰</td><td> 352340 </td><td></td></tr><tr><td>的T25瓶</td><td>文化洁具</td><td> nalge NUNC国际</td><td> 136196 </td><td></td></tr><tr><td> T80瓶</td><td>文化洁具</td><td> nalge NUNC国际</td><td> 178905 </td><td></td></tr><tr><td> 15毫升管</td><td>文化洁具</td><td>屋宇署猎鹰</td><td> 352096 </td><td></td></tr><tr><td> 50毫升管</td><td>文化洁具</td><td>屋宇署猎鹰</td><td> 352070 </td><td&gt; </td></tr><tr><td>表皮生长因子</td><td>生长因子</td><td>研发</td><td> 2028-EG </td><td></td></tr><tr><td> B-FGF </td><td>生长因子</td><td>研发</td><td> 3139-FB： </td><td></td></tr><tr><td>肝素</td><td>生长因子</td><td>西格玛</td><td> H4784 </td><td>重组在PBS </td></tr><tr><td>抗PSA-NCAM的-PE </td><td>抗体</td><td>美天旎生物技术</td><td> 130-093-274 </td><td></td></tr><tr><td> PE标记的鼠IgG1 </td><td>同型对照抗体</td><td>屋宇署Pharminogen </td><td> 556029 </td><td></td></tr><tr><td>肝素</td><td>生长因子</td><td>西格玛</td><td> H4784 </td><td>重组在PBS </td></tr><tr><td> HrBMP-4 </td><td>生长因子</td><td>研发</td><td> 314-BP-010 </td><td></td></tr><tr><td>聚-L-Ornit海因</td><td>试剂</td><td>西格玛</td><td> P4957 </td><td></td></tr><tr><td>胎牛血清</td><td>中等补编</td><td> Gibco公司</td><td> 10099-141 </td><td></td></tr><tr><td> GFAP的</td><td>抗体</td><td> DAKO公司</td><td> Z0334 </td><td></td></tr><tr><td> B - 三微管蛋白</td><td>抗体</td><td> Promega公司</td><td> G7121 </td><td></td></tr></table> *为了使下摆，搭配1的MEM的X 10升包，1M肝素钠和160毫升，并带来8.75 L使用蒸馏水的体积。设置最终的pH值至7.4，并储存在4°C

the neuroblast assay: an assay for the generation and enrichment of neuronal progenitor cells from differentiating neural stem cell progeny using flow cytometry

神经干细胞（NS​​Cs）可以分离和扩大规模大，使用的神经球法和中枢神经系统（CNS）的三个主要类型的细胞分化，即星形胶质细胞，少突胶质细胞和神经元。这些特点使神经干细胞和祖细胞在体外研究，如药物筛选，神经毒理学和电，也为许多神经系统疾病的细胞替代疗法的一个宝贵的可再生的细胞来源。然而，在实践中，国科会的后代，低生产的神经元和少突胶质细胞和星形胶质细胞的分化限制其临床应用后优势的异质性。在这里，我们描述了一个新颖的方法，利用荧光激活细胞分选（FACS）技术，从国科会的后代小鼠未成熟神经元的生成和随后净化。使用这种方法，一个非常丰富的神经祖细胞的人口可达到机智豪特任何明显的星形胶质细胞和真正的国科会污染。该程序包括分化E14小鼠神经节隆起，使用的神经球检测其物理（大小和内部的复杂性）和荧光性能，使用流式细胞仪技术的基础上，由未成熟的神经细胞的分离和富集分离和扩大NSC的后代。总体而言，它需要5-7天，产生神经球和6-8天来区分国科会的后代和分离高纯度未成熟的神经细胞。

Watch this Scientific Journal Video about The Neuroblast Assay: An Assay for the Generation and Enrichment of Neuronal Progenitor Cells from Differentiating Neural Stem Cell Progeny Using Flow Cytomet

The Neuroblast Assay: An Assay for the Generation and Enrichment of Neuronal Progenitor Cells from Differentiating Neural Stem Cell Progeny Using Flow Cytometry

这个视频协议，表明了新方法的产生和后续的纯化，根据自己的身体（大小和内部的粒度）和荧光性能，使用流式细胞仪技术从可再生来源的神经干细胞（NS​​Cs）的神经祖细胞。

神经母细胞含量：从区分神经干细胞后代使用流式细胞仪，神经祖细胞的生成和富集含量

Neural stem cells (NSCs) can be isolated and expanded in large-scale, using the neurosphere assay and differentiated into the three major cell types of ...

Research

JoVE Journal

Neuroscience

Isolation and Expansion of the Adult Mouse Neural Stem Cells Using the Neurosphere Assay

成年鼠神经干细胞的分离和扩大使用神经干含量

Isolation and expansion of the putative neural stem cells (NSCs) from the adult murine brain was first described by Reynolds and Weiss in 1992 employing a ...

科研

神经科学

Establishing Embryonic Mouse Neural Stem Cell Culture Using the Neurosphere Assay

建立小鼠胚胎神经干细胞培养神经球含量

In mammalians, stem cells acts as a source of undifferentiated cells to maintain cell genesis and renewal in different tissues and organs during the life ...

Neural-Colony Forming Cell Assay: An Assay To Discriminate Bona Fide Neural Stem Cells from Neural Progenitor Cells

神经细胞集落形成细胞分析检测，以区分善意的神经祖细胞的神经干细胞

The neurosphere assay (NSA) is one of the most frequently used methods to isolate, expand and also calculate the frequency of neural stem cells (NSCs). ...

Live-cell Imaging of Sensory Organ Precursor Cells in Intact Drosophila Pupae

Live-cell Imaging of Sensory Organ Precursor Cells in Intact Drosophila Pupae

在完整的感觉器官前体细胞的活细胞成像果蝇蛹

Since the discovery of Green Fluorescent Protein (GFP), there has been a revolutionary change in the use of live-cell imaging as a tool for understanding ...

Isolation and Culture of Neural Crest Cells from Embryonic Murine Neural Tube

从小鼠胚胎神经管的神经嵴细胞的分离和培养

The embryonic neural crest (NC) is a multipotent progenitor population that originates at the dorsal aspect of the neural tube, undergoes an epithelial to ...

Feeder-free Derivation of Neural Crest Progenitor Cells from Human Pluripotent Stem Cells

从人多能神经嵴前体细胞的无饲养推导干细胞

Human pluripotent stem cells (hPSCs) have great potential for studying human embryonic development, for modeling human diseases in the dish and as a ...

Flow Cytometry Protocols for Surface and Intracellular Antigen Analyses of Neural Cell Types

神经细胞类型的流式细胞仪协议的表面和细胞内抗原分析

Flow cytometry has been extensively used to define cell populations in immunology, hematology and oncology. Here, we provide a detailed description of ...

Cell Sorting of Neural Stem and Progenitor Cells from the Adult Mouse Subventricular Zone and Live-imaging of their Cell Cycle Dynamics

神经干细胞和祖细胞来自成年小鼠脑室下区细胞分选和实时成像的细胞周期动力学

Neural stem cells (NSCs) in the subventricular zone of the lateral ventricles (SVZ) sustain olfactory neurogenesis throughout life in the mammalian brain. ...

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. ...

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 ...