Manipulation of neutrophil-like HL-60 cells for the study of directed cell migration.
Manipulation of neutrophil-like HL-60 cells for the study of directed cell migration.
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DOI：
10.1007/978-1-60761-404-3_9
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发表时间：
2010
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影响因子：
--
通讯作者：
Weiner, Orion D
中科院分区：
文献类型：
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作者：
Millius, Arthur;Weiner, Orion D
许多细胞在趋化性这一过程中会响应外界信号而进行定向细胞迁移。这种能力对许多单细胞生物的捕食和交配、多细胞生物的发育以及免疫系统的功能都至关重要。由于其相对易于操作且具有强大的趋化能力，类中性粒细胞系（HL - 60）一直是分析定向细胞迁移的有力系统。在本章中，我们描述了HL - 60细胞的维持和瞬时转染，并解释了如何通过两种标准的趋化性检测方法（微吸管法和EZ - TAXIS法）分析它们的行为。最后，我们展示了如何固定和染色极化细胞的肌动蛋白细胞骨架以用于荧光显微镜成像。
Many cells undergo directed cell migration in response to external cues in a process known as chemotaxis. This ability is essential for many single-celled organisms to hunt and mate, the development of multicellular organisms, and the functioning of the immune system. Because of their relative ease of manipulation and their robust chemotactic abilities, the neutrophil-like cell line (HL-60) has been a powerful system to analyze directed cell migration. In this chapter, we describe the maintenance and transient transfection of HL-60 cells and explain how to analyze their behavior with two standard chemotactic assays (micropipette and EZ-TAXIS). Finally, we demonstrate how to fix and stain the actin cytoskeleton of polarized cells for fluorescent microscopy imaging.