Empirical chemosensitivity testing in a spheroid model of ovarian cancer using a microfluidics-based multiplex platform
Empirical chemosensitivity testing in a spheroid model of ovarian cancer using a microfluidics-based multiplex platform
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DOI：
10.1063/1.4774309
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发表时间：
2013-01-01
期刊：
影响因子：
3.2
通讯作者：
Mes-Masson, Anne-Marie
中科院分区：
文献类型：
--
作者：
Das, Tamal;Meunier, Liliane;Mes-Masson, Anne-Marie
使用生物标记物来推断患者的药物反应是积极的，但这种方法的重大挑战，包括复杂的通路相互联系，限制了它的应用。可以说，肿瘤敏感性的直接经验测试将提供更可靠的预测价值，尽管它在很大程度上由于与这种方法相关的技术困难而获得的关注很少。我们假设，最近开发的微技术的应用，与更复杂的三维细胞培养相结合，可以提供一个解决其中一些问题的模型。作为概念的证明，我们开发了一种微流控装置，将浆液性卵巢癌细胞系TOV112D的球体包裹起来，并检测它们对卡铂和紫杉醇的化学反应，这两种药物通常用于卵巢癌的治疗。为了索引化学反应，我们分析了在微流控装置内不同药物浓度和治疗时间下，球体内死亡分数的时空演变，通过生命染料和共聚焦显微镜判断。为了反映微环境的影响，我们测试了外源性细胞外基质和球体形成过程中补充血清对其化疗反应的影响。与单层培养相比，球形细胞显示出更强的抗药性。在球体形成过程中，同时存在细胞外基质和高血清浓度，进一步增加了这种抵抗力。在暴露于化疗药物后，整个球体的细胞死亡情况并不是一致的。球体中心细胞死亡率最高，周边细胞死亡率最低。这些结果证明了该方法的有效性，并为进一步利用微流控技术研究卵巢癌的化疗反应奠定了基础。在未来，这样的微型设备可以提供框架，在适合临床决策的时间范围内进行药物敏感性测试。(C)2013年美国物理研究所。[http://dx.doi.org/10.1063/1.4774309]
The use of biomarkers to infer drug response in patients is being actively pursued, yet significant challenges with this approach, including the complicated interconnection of pathways, have limited its application. Direct empirical testing of tumor sensitivity would arguably provide a more reliable predictive value, although it has garnered little attention largely due to the technical difficulties associated with this approach. We hypothesize that the application of recently developed microtechnologies, coupled to more complex 3-dimensional cell cultures, could provide a model to address some of these issues. As a proof of concept, we developed a microfluidic device where spheroids of the serous epithelial ovarian cancer cell line TOV112D are entrapped and assayed for their chemoresponse to carboplatin and paclitaxel, two therapeutic agents routinely used for the treatment of ovarian cancer. In order to index the chemoresponse, we analyzed the spatiotemporal evolution of the mortality fraction, as judged by vital dyes and confocal microscopy, within spheroids subjected to different drug concentrations and treatment durations inside the microfluidic device. To reflect microenvironment effects, we tested the effect of exogenous extracellular matrix and serum supplementation during spheroid formation on their chemotherapeutic response. Spheroids displayed augmented chemoresistance in comparison to monolayer culturing. This resistance was further increased by the simultaneous presence of both extracellular matrix and high serum concentration during spheroid formation. Following exposure to chemotherapeutics, cell death profiles were not uniform throughout the spheroid. The highest cell death fraction was found at the center of the spheroid and the lowest at the periphery. Collectively, the results demonstrate the validity of the approach, and provide the basis for further investigation of chemotherapeutic responses in ovarian cancer using microfluidics technology. In the future, such microdevices could provide the framework to assay drug sensitivity in a timeframe suitable for clinical decision making. (C) 2013 American Institute of Physics. [http://dx.doi.org/10.1063/1.4774309]