Martinez-Serra Jordi, Gutierrez Antonio, Muñoz-Capó Saúl, Navarro-Palou María, Ros Teresa, Amat Juan Carlos, Lopez Bernardo, Marcus Toni F, Fueyo Laura, Suquia Angela G, Gines Jordi, Rubio Francisco, Ramos Rafael, Besalduch Joan
Department of Hematology, University Hospital Son Espases, Palma de Mallorca, Balearic Islands, Spain.
Department of Clinical Analysis, University Hospital Son Espases, Palma de Mallorca, Balearic Islands, Spain.
Onco Targets Ther. 2014 Jun 12;7:985-94. doi: 10.2147/OTT.S62887. eCollection 2014.
The xCELLigence system is a new technological approach that allows the real-time cell analysis of adherent tumor cells. To date, xCELLigence has not been able to monitor the growth or cytotoxicity of nonadherent cells derived from hematological malignancies. The basis of its technology relies on the use of culture plates with gold microelectrodes located in their base. We have adapted the methodology described by others to xCELLigence, based on the pre-coating of the cell culture surface with specific substrates, some of which are known to facilitate cell adhesion in the extracellular matrix. Pre-coating of the culture plates with fibronectin, compared to laminin, collagen, or gelatin, significantly induced the adhesion of most of the leukemia/lymphoma cells assayed (Jurkat, L1236, KMH2, and K562). With a fibronectin substrate, nonadherent cells deposited in a monolayer configuration, and consequently, the cell growth and viability were robustly monitored. We further demonstrate the feasibility of xCELLigence for the real-time monitoring of the cytotoxic properties of several antineoplastic agents. In order to validate this technology, the data obtained through real-time cell analysis was compared with that obtained from using the 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide method. This provides an excellent label-free tool for the screening of drug efficacy in nonadherent cells and discriminates optimal time points for further molecular analysis of cellular events associated with treatments, reducing both time and costs.
xCELLigence系统是一种新的技术方法，可对贴壁肿瘤细胞进行实时细胞分析。迄今为止，xCELLigence尚无法监测源自血液系统恶性肿瘤的非贴壁细胞的生长或细胞毒性。其技术基础依赖于使用底部带有金微电极的培养板。我们已将其他人描述的方法应用于xCELLigence，该方法基于用特定底物对细胞培养表面进行预包被，其中一些底物已知可促进细胞在细胞外基质中的黏附。与层粘连蛋白、胶原蛋白或明胶相比，用纤连蛋白预包被培养板可显著诱导所检测的大多数白血病/淋巴瘤细胞（Jurkat、L1236、KMH2和K562）的黏附。使用纤连蛋白底物时，非贴壁细胞以单层形式沉积，因此能够有力地监测细胞生长和活力。我们进一步证明了xCELLigence用于实时监测几种抗肿瘤药物细胞毒性特性的可行性。为了验证该技术，将通过实时细胞分析获得的数据与使用3-(4,5-二甲基噻唑-2-基)-2,5-二苯基四氮唑溴盐法获得的数据进行了比较。这为筛选非贴壁细胞中的药物疗效提供了一种出色的无标记工具，并可区分与治疗相关的细胞事件进行进一步分子分析的最佳时间点，从而减少时间和成本。
