Name: generation of neural stem cells from discarded human fetal cortical tissue
Uploaded: 2011-05-25T12:00:00
Description: Watch this Scientific Journal Video about Generation of Neural Stem Cells from Discarded Human Fetal Cortical Tissue at JoVE.com

这项工作是支持部分由美国国立卫生研究院：HD054347和NS063997 - 01到VLS的。通过国务院卫生合同＃C024324到VLS的纽约帝国州干细胞基金中的一部分，这项工作也得到了支持。这里所表达的意见仅为作者个人的，不一定反映帝国干细胞，纽约州卫生署，纽约州的。 VLS的是一个多丽丝公爵临床科学家发展奖得主。我们也感谢他的反O1，反物理学抗体的礼物教授蒂莫西Vartanian。

1。神经干细胞培养的清扫和维护的解决方案和材料的制备<ol><li>提前准备100毫升夹层中（KNOCKOUT的DMEM/F12，Invitogen）和冷藏。 </li><li> Prepare100毫升培养液（干临NSC可持续森林管理，Invitrogen公司），并保持在37 ° C在水浴。 </li><li>准备长期长期冷冻保存的细胞的细胞冷冻培养基（KNOCKOUT的DMEM/F12 +10％胎牛血清+ 5％DMSO）。 </li><li>如果需要，准备为4％多聚甲醛（PFA）组织固定。 </li><li>消毒，灭菌镊子...

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是对文化的新鲜组织和人类细胞系生产的各种方法。从历史上看，已经收获和新鲜组织培养，立即在中枢神经系统中产生的各种细胞类型。然而，这种做法显然是有限的，可这在人类样本的情况下，通常是相当小的样本数量。鉴于操纵的最小程度，新鲜培养的神经细胞提供最可靠的实验系统，通过限制延长培养潜在的工件。供应有限导致的第二种方法产生人体细胞线，已通过插入各种癌基因12...

<table border="1"><tbody><tr><th>试剂名称</th><th>公司</th><th>目录编号</th><th>评论（可选） </th></tr><tr><td> KNOCKOUT的DMEM/F12 </td><td> Invitrogen公司</td><td> 12660-012 </td><td>解离中等</td></tr><tr><td>干临国科会可持续森林管理</td><td> Invitrogen公司</td><td> A10509 - 01 </td><td>培养基</td></tr><tr><td>胎牛血清</td><td> Invitrogen公司</td><td> 10091-148 </td><td>冷冻介质</td></tr><tr><td> Hanks液（ 钙，镁2） </td><td> Invitrogen公司</td><td> 14175-095 </td><td>解离中等</td></tr><tr><td>二甲基亚砜</td><td> Sigma - Aldrich公司</td><td> D2650 </td><td>冷冻介质</td></tr><tr><td> EDTA </td><td> Sigma - Aldrich公司</td><td> 431788 </td><td>解离中等</td></tr><tr><td>多聚甲醛</td><td> Sigma - Aldrich公司</td><td> 158127 </td><td>固定的解决方案</td></tr><tr><td>碱性成纤维细胞生长因子</td><td>研发</td><td> 234 - FSE </td><td>分化培养基</td></tr><tr><td> SHH </td><td>研发</td><td> 1845 - SH </td><td>分化培养基</td></tr><tr><td> PDGF - AA </td><td>研发</td><td> 221 - AA </td><td>分化培养基</td></tr><tr><td> B27 </td><td> Invitrogen公司</td><td> 17504-044 </td><td>分化培养基</td></tr><tr><td>鼠抗- MAP2 </td><td> Sigma - Aldrich公司</td><td> M2320 </td><td> 1:200 </td></tr><tr><td>兔抗- DCX </td><td>细胞信号转导</td><td> 4604s </td><td> 1:200 </td></tr><tr><td>兔抗GFAP </td><td> DAKO公司</td><td> Z0334 </td><td> 1:200 </td></tr><tr><td>兔抗- S100B </td><td> DAKO公司</td><td> Z0311 </td><td> 1:200 </td></tr><tr><td>兔抗O1群</td><td>教授Timothy Vartanian礼物* </td><td></td><td> 1:50 </td></tr><tr><td>兔抗- O4 </td><td>教授Timothy Vartanian礼品* </td><td></td><td> 1:50 </td></tr><tr><td> 40微米细胞过滤器</td><td>屋宇署猎鹰</td><td> 352340 </td><td></td></tr></tbody></table> *提摩太Vartanian，医学博士，哲学博士，美国威尔康乃尔医学院神经病学和神经科学，纽约，系

generation of neural stem cells from discarded human fetal cortical tissue

神经干细胞（NS​​Cs）居住在沿脑室区神经上皮的皮质板的发展。这些早期的祖细胞，最终上升到中间的祖先，后来，大脑皮层的各种神经元和神经胶质细胞亚型。发电量和扩大人类神经干细胞从废弃的正常胎儿组织（所谓的神经球）提供了一种手段，用以直接研究人体正常的国科会发展 1-5的功能方面。这种方法也可以直接向从已知的神经系统疾病的神经干细胞的生成，从而给予的机会，以确定疾病的进程，改变祖细胞的增殖，迁移和分化6-9。我们一直专注于识别人类唐氏综合征的神经干细胞可能有助于加速老年痴呆症的疾病表型10,11的病理机制。无论在体内还是在体外培养的小鼠模型，可以复制的基因位于人类21号染色体上的相同的剧目。 这里我们使用一个简单而可靠的的方法隔离综合征从流产的人类胚胎皮层神经干细胞，并在文化中成长。该方法提供有限的解剖标志，细胞分选，电镀和人类神经干细胞传代收获组织，解剖的具体方面。我们还提供一些基本的协议更有选择性的细胞亚型的人类神经干细胞诱导分化。

Watch this Scientific Journal Video about Generation of Neural Stem Cells from Discarded Human Fetal Cortical Tissue at JoVE.com

Generation of Neural Stem Cells from Discarded Human Fetal Cortical Tissue

描述一个简单和可靠的方法是从丢弃的人类胚胎皮层组织的神经干细胞的分离和培养。从已知的人类神经系统疾病的病理细胞和分子过程的特性，可用于文化，以及提供一个平台，以评估药理药效。

从废弃的人类胎儿大脑皮质组织神经干细胞的生成

Neural stem cells (NSCs) reside along the ventricular zone neuroepithelium during the development of the cortical plate. These early progenitors ...

Research

JoVE Journal

Neuroscience

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). ...

科研

神经科学

Isolation and Culture of Neural Crest Stem Cells from Human Hair Follicles

从人类毛囊神经嵴干细胞的分离和培养的

Hair follicles undergo lifelong growth and hair cycle is a well-controlled process involving stem cell proliferation and quiescence. Hair bulge is a ...

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 ...

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 ...

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 ...

A cGMP-applicable Expansion Method for Aggregates of Human Neural Stem and Progenitor Cells Derived From Pluripotent Stem Cells or Fetal Brain Tissue

人类神经的聚集体cGMP的适用展开法干细胞和祖细胞来源从多能干细胞或胎儿脑组织

A cell expansion technique to amass large numbers of cells from a single specimen for research experiments and clinical trials would greatly benefit the ...

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 ...

Modeling Astrocytoma Pathogenesis In Vitro and In Vivo Using Cortical Astrocytes or Neural Stem Cells from Conditional, Genetically Engineered Mice

Modeling Astrocytoma Pathogenesis In Vitro and In Vivo Using Cortical Astrocytes or Neural Stem Cells from Conditional, Genetically Engineered Mice

建模星形细胞瘤发病机制<em&gt;在体外</em&gt;和<em&gt;体内</em&gt;使用星形胶质细胞或条件，遗传工程小鼠神经干细胞

Current astrocytoma models are limited in their ability to define the roles of oncogenic mutations in specific brain cell types during disease ...

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 ...

Development of a Refined Protocol for Trans-scleral Subretinal Transplantation of Human Retinal Pigment Epithelial Cells into Rat Eyes

人视网膜色素上皮细胞向大鼠眼巩膜视网膜移植精制方案的研制

Degenerative retinal diseases such as age-related macular degeneration (AMD) are the leading cause of irreversible vision loss worldwide. AMD is ...