A real time chemotaxis assay unveils unique migratory profiles amongst different primary murine macrophages.
A real time chemotaxis assay unveils unique migratory profiles amongst different primary murine macrophages.
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DOI：
10.1371/journal.pone.0058744
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Greaves DR
中科院分区：
文献类型：
--
作者：
Iqbal AJ;Regan-Komito D;Christou I;White GE;McNeill E;Kenyon A;Taylor L;Kapellos TS;Fisher EA;Channon KM;Greaves DR
趋化性测定是用于研究炎症介质如CC趋化因子的生物活性的宝贵工具，CC趋化因子已涉及广泛的慢性炎症性疾病。传统的趋化系统，如改良的Boyden室，在捕获的数据方面是有限的，因为在单个时间点分析测定。我们报告的优化和验证的无标记，实时细胞迁移试验的基础上，电细胞阻抗来测量不同的原代小鼠巨噬细胞群体的趋化性，在一系列的CC趋化因子和其他化学引诱物信号分子。我们清楚地证明了不同鼠巨噬细胞群体迁移行为的关键差异，并表明这种动态系统测量的是真正的巨噬细胞趋化性，而不是趋化性或趋避性。我们强调了Gαi信号和肌动蛋白细胞骨架重排的绝对要求，正如百日咳毒素和细胞松弛素D抑制所证明的那样。我们还研究了CD14+人单核细胞的趋化性，并证明了不同的单核细胞供体CCL2之间不同的趋化性。这种实时趋化性测定将允许对调节巨噬细胞对趋化因子细胞因子和炎症介质的反应的因素进行详细分析。
Chemotaxis assays are an invaluable tool for studying the biological activity of inflammatory mediators such as CC chemokines, which have been implicated in a wide range of chronic inflammatory diseases. Conventional chemotaxis systems such as the modified Boyden chamber are limited in terms of the data captured given that the assays are analysed at a single time-point. We report the optimisation and validation of a label-free, real-time cell migration assay based on electrical cell impedance to measure chemotaxis of different primary murine macrophage populations in response to a range of CC chemokines and other chemoattractant signalling molecules. We clearly demonstrate key differences in the migratory behavior of different murine macrophage populations and show that this dynamic system measures true macrophage chemotaxis rather than chemokinesis or fugetaxis. We highlight an absolute requirement for Gαi signaling and actin cytoskeletal rearrangement as demonstrated by Pertussis toxin and cytochalasin D inhibition. We also studied the chemotaxis of CD14+ human monocytes and demonstrate distinct chemotactic profiles amongst different monocyte donors to CCL2. This real-time chemotaxis assay will allow a detailed analysis of factors that regulate macrophage responses to chemoattractant cytokines and inflammatory mediators.
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DOI：
10.1083/jcb.146.4.831
发表时间：
1999-08-23
期刊：
The Journal of cell biology
影响因子：
--
作者：
Correia I;Chu D;Chou YH;Goldman RD;Matsudaira P
通讯作者：
Matsudaira P
影响因子：
30.5
作者：
通讯作者：
--
影响因子：
2.9
作者：
Sun, Mingjiao;Fu, Huiying;Ke, Yuehai
通讯作者：
Ke, Yuehai
影响因子：
15.3
作者：
Cash, Jenna L.;Hart, Rosie;Russ, Andreas;Dixon, John P. C.;Colledge, William H.;Doran, Joanne;Hendrick, Alan G.;Carlton, Mark B. L.;Greaves, David R.
通讯作者：
Greaves, David R.
影响因子：
4.4
作者：
Dabbagh, K;Xiao, Y;Lewis, DB
通讯作者：
Lewis, DB