| In vitro Migration Assays for Neural Stem Cells, Intermediate Neurogenic Progenitors and Immature Neurons
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Author:
Date:
2015-01-05
[Abstract] In the vertebrate central nervous system (CNS), different neural precursor populations such as neural stem cells (NSCs), intermediate neurogenic progenitors (INPs) and immature neurons have to migrate from their places of birth to their location of function. Coordinated migration is mediated by direct cell-cell interactions and by extracellular matrix components, chemoattractants as well as repellents. The migration potential of such populations as well as the responsiveness to chemoattractive compounds can be addressed in isolated cells using in vitro migration assays. Here we describe two migration assays, a matrigel migration assay and a Boyden chamber migration assay, which allow the in vitro assessment of neural migration under defined conditions (Ladewig, Koch and ...
[摘要] 在脊椎动物中枢神经系统（CNS）中，不同的神经前体细胞如神经干细胞（NSCs），中间神经源性祖细胞（INPs）和不成熟的神经元必须从他们的出生地迁移到它们的功能位置。配位迁移通过直接的细胞 - 细胞相互作用和细胞外基质组分，化学引诱物以及驱避剂介导。这些群体的迁移潜力以及对化学吸引化合物的反应性可以使用体外迁移测定在分离的细胞中解决。在这里我们描述两个迁移测定，基质胶迁移测定和博伊登室迁移测定，其允许在确定的条件下的神经迁移的体外评估（Ladewig，Koch和Brüstle，2014）。基质胶基质是可溶性基底膜提取物。基质胶基质的主要成分是胶原，层粘连蛋白和蛋白聚糖，它们为迁移细胞提供了底物。在基质胶试验中，可以使用相差显微镜分析迁移。 Boyden室测定法（Richards和McCullough，1984）基于微化学趋化室，其由通过具有限定孔径的膜分开的两个室组成。细胞可以接种在上室中并允许通过孔迁移到下室，其中加载了潜在的趋化剂。可以在固定和免疫组织化学染色后分析细胞迁移。原则上，所描述的方案应该适用于其它细胞群体，例如内皮细胞或癌细胞，使用适合于特定细胞类型的个体需要的条件。
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| Transwell Cell Migration Assay Using Human Breast Epithelial Cancer Cell
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Author:
Date:
2012-02-20
[Abstract] Transwell migration assays have been widely used for studying the motility of different types of cells including metastatic cancer cells. The assay is also useful in screens for compounds that act as chemoattractants or inhibitors of chemotaxis for cells. The assay employs a permeable layer of support, usually a tissue-culture-treated microporous membrane, which is positioned between two compartments that mimic two different sets of microenvironments for cell survival/growth. Cells on one side of the membrane, when sensing chemoattractants placed on the other side of the compartment that diffuses through the membrane, can migrate through the pores in the membrane towards the source of the chemoattractants. Cells that migrate across the membrane can be quantified by fixing and counting. ...
[摘要] Transwell迁移测定已经广泛用于研究不同类型的细胞（包括转移性癌细胞）的运动性。该测定法还可用于筛选充当化学趋化剂或细胞趋化性抑制剂的化合物。该测定使用可渗透的支持层，通常是组织培养处理的微孔膜，其位于模拟用于细胞存活/生长的两种不同组的微环境的两个区室之间。膜的一侧上的细胞，当感测放置在通过膜扩散的隔室的另一侧上的化学引诱物时，可以通过膜中的孔向化学引诱物的来源迁移。穿过膜迁移的细胞可以通过固定和计数来定量。人乳腺上皮腺癌MD-231细胞生长相对较快并且是转移性的。在此使用MB-231细胞系来描述使用transwell装置的体外细胞迁移测定的程序。
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| Tumor Cell Invasion Assay
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Author:
Date:
2012-02-05
[Abstract] Cell invasion assays have been used to study the interactions between tumor cells and the extracellular matrix (ECM), which not only provides a structural scaffold for the cell, but also contains various biological factors for the survival and growth of the cell. ECM gel contains the basic components of the ECM that provides a structural support for the cell to grow and move. Cells can secrete enzymes that degrade certain components of the ECM to move towards chemoattractants, or to simply establish niches for growth. Metastatic tumor cells often show more invasiveness to the ECM gel due to their higher motility and/or enzymatic activity for degrading ECM components. This protocol describes a tumor cell invasion assay to study the interactions between tumor cells and the ECM.
[摘要] 细胞侵袭测定已经用于研究肿瘤细胞和细胞外基质（ECM）之间的相互作用，其不仅为细胞提供结构支架，而且还包含用于细胞存活和生长的各种生物因子。 ECM凝胶含有ECM的基本组分，为细胞生长和移动提供结构支持。 细胞可分泌降解ECM的某些组分以趋向化学引诱物的酶，或简单地建立用于生长的壁龛。 转移性肿瘤细胞通常对ECM凝胶显示更多的侵袭性，这是由于它们对于降解ECM组分的较高的运动性和/或酶活性。 该协议描述了肿瘤细胞侵袭实验，以研究肿瘤细胞和ECM之间的相互作用。
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