Name: identification and isolation of slow-dividing cells in human glioblastoma using carboxy fluorescein succinimidyl ester (cfse)
Uploaded: 2012-04-29T14:00:00
Description: Watch this Scientific Journal Video about Identification and Isolation of Slow-Dividing Cells in Human Glioblastoma Using Carboxy Fluorescein Succinimidyl Ester CFSE at JoVE.com

这项工作是由佛罗里达州，普雷斯顿A.富国小中心的脑肿瘤治疗和美国国立卫生研究院（1R21CA141020-01）脑肿瘤研究中心的支持。

1。准备胶质母细胞瘤单细胞悬液<ol><li>使用的神经球的吸附试验（NSA）如先前所述2,11,12 gliomasphere文化建立和维护。 </li><li>在适当的时间用于传代gliomaspheres的，介质中含有球体除去，放置在一个适当大小的无菌组织培养管中，在室温下5分钟（110克）在800 rpm的转速下离心分离。 </li><li>弃去上清液，和球形的颗粒再悬浮于1毫升的0.05％胰蛋白酶-EDTA和孵育在37℃下在水浴中3-5分钟。 </li><li>...

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越来越多的研究已经证明一个缓慢的分割的子隔室的细胞在癌症有助于肿瘤的形成和治疗抵抗2-8的重要性。因此，这是至关重要的描述和标准化的实验方法，使此特定的细胞或更普遍的亚群的研究比较亚群具有不同的生长速率。使用的CFSE识别和分离部分的细胞根据其细胞分裂的速度是一个起点，进一步描述这些特定的细胞组分，有助于推进我们的理解，在肿瘤的异质性。在这个协议中，?...

<table border="1"><tbody><tr><td> 的试剂的名称 </td><td> 类型 </td><td> 公司 </td><td> 目录编号 </td></tr><tr><td> NeuroCult NSC基础培养基（人） </td><td>中</td><td>干细胞技术</td><td> 05750 </td></tr><tr><td> NeuroCult补充NSC增殖（人） </td><td>中补充</td><td>干细胞技术</td><td> 05753 </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></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></tr><tr><td>青霉素/链霉素</td><td>试剂</td><td> Gibco公司</td><td> 15140-122 </td></tr><tr><td> T25烧瓶</td><td>培养器皿</td><td> Nalge Nunc公司国际</td><td> 136196 </td></tr><tr><td> T80烧瓶</td><td>培养器皿</td><td> Nalge Nunc公司国际</td><td> 178905 </td></tr><tr><td> 15 mL管</td><td>培养器皿</td><td> BD猎鹰</td><td> 352096 </td></tr><tr><td> 50ml试管</td><td>培养器皿</td><td> BD猎鹰</td><td> 352070 </td></tr><tr><td> EGF </td><td>生长因子</td><td> R＆D </td><td> 2028-EG </td></tr><tr><td> CellTrace CFSE细胞增殖试剂盒</td><td>跟踪染料荧光细胞分裂</td><td> Molecular Probes公司，Invitrogen公司</td><td> C34554 </td></tr></tbody></table>

identification and isolation of slow-dividing cells in human glioblastoma using carboxy fluorescein succinimidyl ester (cfse)

肿瘤的异质性表示支持肿瘤的鲁棒性的一个基本特征，并提出了一种中央障碍的发展的治疗策略1。为了克服肿瘤的异质性的问题，它是必不可少的表型，（EPI）的基因和功能的识别和鉴定的肿瘤亚群，推动特定疾病的病理学和代表临床相关指标的检测和工具，使开发。这是目前公认的肿瘤表现出不同的子细胞与细胞分裂的不同频率的分数，慢骑自行车的功能标准与多种癌症，包括脑癌，乳腺癌，皮肤肿瘤形成能力呈正相关胰腺以及白血病2-8。的荧光染料羧基荧光素琥珀酰亚胺基酯（CFSE）已被用于在体外和体内细胞的分裂频率跟踪在血源性吨umors和实体肿瘤如神经母细胞瘤2,7,8。细胞穿透物的非荧光的CFSE的前体药物通过细胞内酯酶转换成荧光的化合物，它被保留在细胞内，由共价结合的蛋白质，通过反应的琥珀酰亚胺基团与细胞内的胺基团，以形成稳定的酰胺键9。荧光染料同样分布之间的子细胞时分裂，导致细胞分裂的荧光强度与每个减半。这使得追踪细胞周期的频率高达八至十个师10轮。用CFSE保留容量与脑肿瘤细胞的识别和分离证明富集在肿瘤干细胞的活性2缓慢循环亚群（顶部5％的染料保持细胞）。 该协议描述了技术CFSE染色细胞内的个别人群的隔离一个的人glio文化母细胞瘤（GBM）衍生的细胞具有不同的部门使用流式细胞仪2。该技术一直凭借自己的能力，保留CFSE标记的识别和分离脑肿瘤慢骑自行车的细胞群。

Watch this Scientific Journal Video about Identification and Isolation of Slow-Dividing Cells in Human Glioblastoma Using Carboxy Fluorescein Succinimidyl Ester CFSE at JoVE.com

Identification and Isolation of Slow-Dividing Cells in Human Glioblastoma Using Carboxy Fluorescein Succinimidyl Ester CFSE

此视频协议演示的荧光染料羧基荧光素琥珀酰亚胺基酯（CFSE）的应用程序的不同的子种群的细胞分裂的频率的基础上在人成胶质细胞瘤细胞的识别和分离。

慢分裂的细胞在人脑胶质瘤羧基荧光素琥珀酰亚胺酯（CFSE）的分离与鉴定

Tumor heterogeneity represents a fundamental feature supporting tumor robustness and presents a central obstacle to the development of therapeutic ...

Research

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Identification and Isolation of Slow-Dividing Cells in Human Glioblastoma Using Carboxy Fluorescein Succinimidyl Ester (CFSE)

Isolation, Characterization and Comparative Differentiation of Human Dental Pulp Stem Cells Derived from Permanent Teeth by Using Two Different Methods

人牙髓干细胞的分离，鉴定和比较性恒牙使用两种不同的方法，得出

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