Applications of stripe assay in the study of CXCL12-mediated neural progenitor cell migration and polarization.
Applications of stripe assay in the study of CXCL12-mediated neural progenitor cell migration and polarization.
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DOI：
10.1016/j.biomaterials.2015.08.052
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发表时间：
2015-12
期刊：
影响因子：
14
通讯作者：
Zheng JC
中科院分区：
文献类型：
--
作者：
Zhang M;Song A;Lai S;Qiu L;Huang Y;Chen Q;Zhu B;Xu D;Zheng JC
神经前体细胞（NPC）的极化和迁移对于胚胎脑发育和脑损伤后的神经发生至关重要。尽管基质衍生因子-1 α（SDF-1α，CXCL 12）及其受体CXCR 4在发育中的脑中介导NPC的迁移，但其动态细胞过程和结构相关的分子事件仍然是未知的。基于Transwell和微流体的测定是有效研究细胞迁移的经典测定。然而，这两种方法在分析单个细胞时都有局限性。在这项研究中，我们修改了条纹试验，并扩大其在NPC极化和细胞内分子事件与CXCL 12介导的迁移的研究中的应用。响应于局部CXCL 12，NPC在条纹测定中形成板状伪足。此外，CXCR 4和Rac 1迅速重新分布到板状伪足区域，表明它们在CXCL 12刺激后在NPC极化中的作用。尽管该测定中的趋化因子条带提供了可以最好地用于通过免疫细胞化学研究细胞极化和迁移的浓度梯度，但它们也可以生成具有可比质量的实时成像数据。条纹法是研究细胞运动及其相关分子机制的一种直观、动态、经济的方法。
The polarization and migration of neural progenitor cells (NPCs) are critical for embryonic brain development and neurogenesis after brain injury. Although stromal-derived factor-1α (SDF-1α, CXCL12) and its receptor CXCR4 are well-known to mediate the migration of NPCs in the developing brain, the dynamic cellular processes and structure-related molecular events remain elusive. Transwell and microfluidic-based assays are classical assays to effectively study cellular migration. However, both of them have limitations in the analysis of a single cell. In this study, we modified the stripe assay and extended its applications in the study of NPC polarization and intracellular molecular events associated with CXCL12-mediated migration. In response to localized CXCL12, NPCs formed lamellipodia in the stripe assay. Furthermore, CXCR4 and Rac1 quickly re-distributed to the area of lamellipodia, indicating their roles in NPC polarization upon CXCL12 stimulation. Although the chemokine stripes in the assay provided concentration gradients that can be best used to study cellular polarization and migration through immunocytochemistry, they can also generate live imaging data with comparable quality. In conclusion, stripe assay is a visual, dynamic and economical tool to study cellular mobility and its related molecule mechanisms.