基于一个基质胶管形成实验，以评估对肿瘤细胞的血管生成活性
Summary
管形成实验是用来评估肿瘤细胞的血管活性。
Abstract
在过去的几十年里，一个管形成实验使用的增长因素，减少基质胶通常已经证明在 体外培养1-5的血管内皮细胞的血管生成活性。然而，最近越来越多的证据表明，这种检测不仅限于测试对血管内皮细胞的行为。相反，它也被用来测试的肿瘤细胞数量的能力，培养血管表型 6-8 。这种能力是与他 们的血管生成xenotransplanted动物，一个过程被称为血管生成拟态（VM）的9所确定的行为是一致的。有大量的证据表明，肿瘤细胞介导的虚拟机中起着至关重要的作用，在肿瘤的发展，独立于血管内皮细胞血管生成6， 10-13。例如，肿瘤细胞被发现参与组织样本中的血液灌流，血管通道形成黑色素瘤和胶质母细胞瘤患者，8，10，11 。在这里，我们描述了这种管状网络检测，作为一个有用的工具，在评价肿瘤细胞的血管生成活性。我们发现，一些肿瘤细胞株，如黑色素瘤B16F1细胞，胶质母细胞瘤U87细胞和乳腺癌MDA – MB – 435细胞能够形成血管管;但也有一些如结肠癌HCT116细胞。此外，这种血管表型是依赖于基质胶上镀的手机号码。因此，这种检测可以作为强大的工具，屏幕上的多种细胞类型，包括血管细胞，肿瘤细胞以及其它细胞的血管潜力。
Protocol
1。基于一个基质胶管形成实验，以评估对肿瘤细胞的血管生成活性所有的肿瘤细胞和微血管内皮细胞的制备：U87细胞，黑色素瘤细胞，乳腺癌MDA – MB – 435，和结肠癌细胞HCT116 B16F1如脑肿瘤细胞生长，补充10％胎牛血清和青霉素/链霉素的DMEM（Invitrogen公司）。内皮细胞株人微血管（HMVECs）内皮细胞培养EBM2媒介为辅1微克/毫升氢化可的松和1 ng / ml的表皮生长因子，10％胎牛血清和青霉素/链霉素…
Discussion
为了这个实验取得成功，基质胶的质量，应先进行测试。一个小样本，可从屋宇署生物科学，预先使用HMVECs运行检测。不同批次的产品可能会显示不同的素质，其中一些地段不提供管形成的最佳条件。第二，任何气泡应避免在96孔板中装入基质胶等份，因为泡沫可以破坏肾小管形成。如果发现在一个良好的微小气泡，基质胶可以立即移动返回到原来的基质胶管在实验过程中应保持在冰上。此外，?…
Declarações
The authors have nothing to disclose.
Acknowledgements
这项工作是由NCI R01 CA120659（RS）的支持。
Materials
| Name of the reagent | Company | Catalogue number |
|---|---|---|
| DMEM | Invitrogen | 11995 |
| FBS | Invitrogen | 16000-044 |
| Growth factor-reduced Matrigel | BD Bioscience | 47743-720 |
| EBM2 kit | Lonza | CC-3156 |
| Nikon ECLIPSE TS100 microscope | Nikon |
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Citar este artigo
Francescone III, R. A., Faibish, M., Shao, R. A Matrigel-Based Tube Formation Assay to Assess the Vasculogenic Activity of Tumor Cells. J. Vis. Exp. (55), e3040, doi:10.3791/3040 (2011).