Mazharian Alexandra
Centre for Cardiovascular Sciences, Institute of Biomedical Research, School of Clinical and Experimental Medicine, College of Medical and Dental Sciences, University of Birmingham, Birmingham, UK.
Methods Mol Biol. 2012;788:275-88. doi: 10.1007/978-1-61779-307-3_19.
Cell migration is a highly integrated multistep process that plays an essential role during development and disease. Megakaryocytes (MKs) are specialized precursor cells that produce platelets and release them into the circulation. MK migration from the proliferative osteoblastic niche within the bone marrow (BM) environment to the capillary-rich vascular niche is an essential step for platelet production. Among the chemokines that may play a central role in cell migration, the stromal cell-derived factor 1α (SDF1α) also known as CXCL12 has been described to act as a potent chemoattractant for MKs. This biological effect is mediated by the SDF1α receptor CXCR4 (Fusin), which is expressed on haematopoietic stem cells, MKs and platelets. The Dunn chemotaxis chamber in conjunction with the time-lapse microscopy is a powerful tool that enables the user to observe directly the morphological response of cells to chemoattractant in real time. This chapter describes the Dunn chemotaxis chamber to study the migration of primary BM-derived MKs in response to a gradient of SDF1α. In combination with genetically modified mice, this provides a powerful approach to directly investigate the role of specific proteins in MK migration and chemotaxis.
细胞迁移是一个高度整合的多步骤过程，在发育和疾病过程中发挥着重要作用。巨核细胞（MKs）是产生血小板并将其释放到循环系统中的特殊前体细胞。MK从骨髓（BM）环境中增殖性成骨细胞龛迁移到富含毛细血管的血管龛是血小板生成的关键步骤。在可能在细胞迁移中起核心作用的趋化因子中，基质细胞衍生因子1α（SDF1α，也称为CXCL12）已被描述为对MKs有强大趋化作用的因子。这种生物学效应由SDF1α受体CXCR4（融合素）介导，CXCR4在造血干细胞、MKs和血小板上表达。Dunn趋化室结合延时显微镜是一种强大的工具，可让使用者实时直接观察细胞对趋化因子的形态学反应。本章介绍使用Dunn趋化室研究原代BM来源的MKs对SDF1α梯度的迁移反应。结合基因改造小鼠，这为直接研究特定蛋白质在MK迁移和趋化中的作用提供了有力方法。
