Comparative analysis of dynamic cell viability, migration and invasion assessments by novel real-time technology and classic endpoint assays.
Comparative analysis of dynamic cell viability, migration and invasion assessments by novel real-time technology and classic endpoint assays.
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DOI：
10.1371/journal.pone.0046536
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发表时间：
2012
期刊：
影响因子：
3.7
通讯作者：
Pauwels P
中科院分区：
文献类型：
--
作者：
Limame R;Wouters A;Pauwels B;Fransen E;Peeters M;Lardon F;De Wever O;Pauwels P
细胞的存活和运动包括无处不在的机制，涉及包括癌症在内的各种(病理)生物过程。我们报告了新型基于阻抗的xCELigence实时细胞分析检测平台与传统的基于标记的终点方法的技术对比分析，从而揭示了高度标准化的实验条件下细胞增殖、细胞毒性、迁移和侵袭的性能特征和相关的动态观察。使用xCELLigence技术对乳腺癌细胞系MDA-MB-231和肺癌细胞系A549的增殖、细胞毒性和迁移进行动态高分辨率评价。在一系列四种细胞浓度下，将增殖动力学与硫代罗丹明B(SRB)分析进行比较，得到了良好的相关性(Spearman‘s Rho 0.688-0.964)。紫杉醇(0-100 nm)的细胞毒作用与SRB(Rho>；0.95)有很好的相关性，IC50值相似。使用Transwell平板进行参考细胞迁移实验，并通过结晶紫染色膜的像素面积计算(Rho 0.90)和提取染料的光密度(OD)测量(Rho>；0.95)进行关联。使用Matrigel包被的Trswells作为标准参照法，观察到单独对MDA-MB-231细胞的侵袭，并通过两种Matrigel密度(Rho>；0.95)的OD读数进行关联。方差成分分析显示，与基于阻抗的迁移和侵袭检测相关的方差增加，这可能是由于这种方法的敏感性造成的。XCELLigence RTCA技术为非侵入性检测细胞活性和运动性提供了一个准确的平台。与传统方法的强相关性意味着对细胞行为和与其他系统的互换性的类似观察，通过仅采用适应的基质表面密度的不同平台上的高度关联的动力学侵袭轮廓来说明。然而，灵敏度的提高意味着标准化的实验条件，以最大限度地减少技术引起的差异。
Cell viability and motility comprise ubiquitous mechanisms involved in a variety of (patho)biological processes including cancer. We report a technical comparative analysis of the novel impedance-based xCELLigence Real-Time Cell Analysis detection platform, with conventional label-based endpoint methods, hereby indicating performance characteristics and correlating dynamic observations of cell proliferation, cytotoxicity, migration and invasion on cancer cells in highly standardized experimental conditions. Dynamic high-resolution assessments of proliferation, cytotoxicity and migration were performed using xCELLigence technology on the MDA-MB-231 (breast cancer) and A549 (lung cancer) cell lines. Proliferation kinetics were compared with the Sulforhodamine B (SRB) assay in a series of four cell concentrations, yielding fair to good correlations (Spearman's Rho 0.688 to 0.964). Cytotoxic action by paclitaxel (0–100 nM) correlated well with SRB (Rho>0.95) with similar IC50 values. Reference cell migration experiments were performed using Transwell plates and correlated by pixel area calculation of crystal violet-stained membranes (Rho 0.90) and optical density (OD) measurement of extracted dye (Rho>0.95). Invasion was observed on MDA-MB-231 cells alone using Matrigel-coated Transwells as standard reference method and correlated by OD reading for two Matrigel densities (Rho>0.95). Variance component analysis revealed increased variances associated with impedance-based detection of migration and invasion, potentially caused by the sensitive nature of this method. The xCELLigence RTCA technology provides an accurate platform for non-invasive detection of cell viability and motility. The strong correlations with conventional methods imply a similar observation of cell behavior and interchangeability with other systems, illustrated by the highly correlating kinetic invasion profiles on different platforms applying only adapted matrix surface densities. The increased sensitivity however implies standardized experimental conditions to minimize technical-induced variance.
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影响因子：
0.7
作者：
De Wever, Olivier;Hendrix, An;Bracke, Marc
通讯作者：
Bracke, Marc
影响因子：
3.3
作者：
Huang, J;Bridges, LC;White, JM
通讯作者：
White, JM
影响因子：
8.4
作者：
Pauwels, B;Korst, AEC;Vermorken, JB
通讯作者：
Vermorken, JB
DOI：
10.1007/978-1-61779-108-6_6
发表时间：
2011-01-01
期刊：
MAMMALIAN CELL VIABILITY: METHODS AND PROTOCOLS
影响因子：
--
作者：
Ke, Ning;Wang, Xiaobo;Abassi, Yama A.
通讯作者：
Abassi, Yama A.
影响因子：
14.9
作者：
Romano, Paolo;Manniello, Assunta;Parodi, Barbara
通讯作者：
Parodi, Barbara