| 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 ...
[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 ...
[摘要] 小室迁移实验已被广泛用于研究不同类型包括转移性肿瘤细胞的细胞活力。该实验对检验趋化因子或细胞趋化抑制剂的化合物是有用的。检测依赖于透水层的支持，通常这是组织培养处理的微孔滤膜，它位于两个小室之间，模仿两个不同的微环境对细胞生存/生长。细胞在膜的一侧，从小室的另一侧感应的趋化因子，可以通过微孔膜迁移。被移植的细胞可以用简单的固定/染色的计数方法量化。人类乳腺上皮腺癌MD - 231细胞生长比较快，是转移性的。 以下就以MB - 231细胞细胞株为例描述细胞迁移实验的基本程序。
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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 ...
[摘要] 细胞侵袭实验用来研究肿瘤细胞和胞外基质(ECM)之间的相互作用。ECM不仅为细胞提供了结构支架，同时也包含了许多细胞生存及生长过程中生物功能因子。细胞可以分泌一种酶，该酶能够降解EMC中特定的组分，从而使细胞向化学引诱剂移动，或者直接形成一种生长的状态。EMC胶模仿体内细胞外基质ECM环境，包含了支撑细胞结构的最基本的组分。转移性肿瘤细胞由于其高迁移和/或降解ECM的酶活从而表现出较强的侵袭性。
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