Millius Arthur, Weiner Orion D
Cardiovascular Research Institute and Department of Biochemistry, University of California, San Francisco, CA, USA.
Methods Mol Biol. 2010;591:147-58. doi: 10.1007/978-1-60761-404-3_9.
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.
许多细胞会响应外部信号，通过一种称为趋化作用的过程进行定向细胞迁移。这种能力对于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至关重要。由于嗜中性粒细胞样细胞系（HL-60）相对易于操作且具有强大的趋化能力，它已成为分析定向细胞迁移的有力系统。在本章中，我们描述了HL-60细胞的培养和瞬时转染，并解释了如何通过两种标准趋化分析方法（微量移液器和EZ-TAXIS）来分析其行为。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进行荧光显微镜成像。
