为研究定向细胞迁移而对嗜中性粒细胞样HL-60细胞进行的操作。
Manipulation of neutrophil-like HL-60 cells for the study of directed cell migration.
作者信息
Millius Arthur, Weiner Orion D
机构信息
Cardiovascular Research Institute and Department of Biochemistry, University of California, San Francisco, CA, USA.
出版信息
Methods Mol Biol. 2010;591:147-58. doi: 10.1007/978-1-60761-404-3_9.
Abstract
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.
摘要
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。
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本文引用的文献
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BMC Infect Dis. 2007 Sep 20;7:110. doi: 10.1186/1471-2334-7-110.
2
Novel RUNX1 isoforms determine the fate of acute myeloid leukemia cells by controlling CD56 expression.新型RUNX1异构体通过控制CD56表达决定急性髓系白血病细胞的命运。
Blood. 2007 Sep 15;110(6):2027-33. doi: 10.1182/blood-2007-02-074203. Epub 2007 Apr 12.
3
Control of cell polarity and motility by the PtdIns(3,4,5)P3 phosphatase SHIP1.由磷脂酰肌醇-3,4,5-三磷酸磷酸酶SHIP1调控细胞极性和运动性
Nat Cell Biol. 2007 Jan;9(1):36-44. doi: 10.1038/ncb1515. Epub 2006 Dec 17.
4
PI(3)Kgamma has an important context-dependent role in neutrophil chemokinesis.磷脂酰肌醇-3激酶γ（PI(3)Kγ）在中性粒细胞趋化运动中具有重要的依赖于环境的作用。
Nat Cell Biol. 2007 Jan;9(1):86-91. doi: 10.1038/ncb1517. Epub 2006 Dec 17.
5
Integrin inhibition through Lyn-dependent cross talk from CXCR4 chemokine receptors in normal human CD34+ marrow cells.通过正常人类CD34+骨髓细胞中CXCR4趋化因子受体的Lyn依赖性串扰实现整合素抑制。
Blood. 2006 Jun 1;107(11):4234-9. doi: 10.1182/blood-2005-08-3343. Epub 2006 Feb 7.
6
Cell migration: integrating signals from front to back.细胞迁移：整合从前到后的信号。
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7
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9
Polarization of chemoattractant receptor signaling during neutrophil chemotaxis.中性粒细胞趋化作用过程中趋化因子受体信号的极化
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10
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