基于生物力学和电学方法多维度、量化地筛选肿瘤单细胞

The diversity of cancer species and high heterogeneity of tumor cells are one of the most important challenges in the clinical treatment of cancer in China. Traditional methods for the detection of metastatic tumor cells mostly rely on cancer markers with high specificity, which are lack of sensitivity, difficult to operate and cannot achieve resolution for single-cell detection. The existing microfluidic platforms can only characterize 1-2 specific physiological cellular parameters of a single cell, which cannot accurately identify tumor cells with high heterogeneity. To solve these problems, we propose the platform based on the biomechanical and electrical analysis for single-cell characterization. The platform integrates surface topographies and stiffness gradients for the guidance of cell migration, micropost sensing arrays for cell traction force measurement, microvortex cavity for cell separation and interdigital electrodes for impedance sensing. Hence, migration characteristics, traction force, size, stiffness and equivalent impedance of different cancer cells could be characterized. Moreover, the platform utilizes radio frequency resonant circuit to transmit the data wirelessly, which helps to realize the modularization of sensing chips and real-time data analysis. The proposed platform utilizes multi-parametric cell characterizations to quantitatively screen different metastatic cancer cells. As the proposed platform is label-free, rapid and convenient, it has great potential in clinical applications, including tumor detection, individualized medical treatment, efficacy evaluation and tumor prognosis monitoring.

癌症种类的多样性与肿瘤细胞的高异质性是我国癌症临床治疗的首要挑战之一。传统方法检测转移肿瘤细胞大多依赖细胞的高特异性标记物，存在敏感度低、操作繁琐和难以实现单细胞检测的问题。而现有基于微流控的细胞检测平台，仅可用于单个细胞的1-2个特定的物理和生理特征的测量，难以准确定型具有高异质性的肿瘤细胞。针对以上问题，本项目基于生物力学和电学分析，制作单细胞检测芯片，利用表面拓扑/硬度结构对细胞迁移的导向作用、微柱体传感器的牵引力测量、微涡旋腔体的流体力分选与插指电极的电阻感应，表征不同癌细胞的迁移特性、牵引力、细胞大小、硬度和等效阻抗，从多维度去量化筛选高特异性的癌细胞。同时，芯片应用电容-电感的射频无线技术，方便实现模块化与实时数据传输。该方案综合了各种结构设计的优势，从多维度准确、快速和方便地量化不同种类的肿瘤，在肿瘤检测、个体化医疗、疗效评估和肿瘤预后监测等方面上有着巨大潜力。

肿瘤细胞的高异质性带来的细胞检测和后续癌症治疗是我国的重大挑战之一。传统方法应用荧光方式检测细胞的高特异性标记物，存在敏感度低、操作繁琐和难以实时监测的问题。.本项目搭建了生物电学、力学分析与培养平台，并且与微流控结合，进行了细胞捕获、培养和不同药物作用的细胞分析。在细胞电学传感方面，我们集成了微流控细胞捕获阵列和电容-电感无线无源电学传感阵列，捕获并且分选了H1299非小细胞肺癌细胞。在细胞的力学分析方面，我们开发了微流控柱形捕获芯片，利用流体力对于不同大小细胞的作用，将不同尺寸大小的癌细胞A549和H446从白细胞中捕获与分离。同时，我们利用了三维（3D）拓扑结构培养细胞球，并且探究了3D和2D培养的人肝癌细胞对于药物浓度的响应。除了细胞检测，我们也拓展了电学检测平台，通过免疫修饰检测特异性的肿瘤蛋白，包括CEA 、AFP和转铁蛋白等。.总结来说，我们已经搭建完成了项目计划中的电学、力学分析平台与拓扑作用下的细胞培养平台。并且，我们集成了微流控浓度梯度结构，分析在不同药物作用下细胞状态的变化。因此，该项目中构建的这些平台可以从各个维度、量化地评估癌细胞的物理、生理变化，在后续的临床检测，药物筛选，个体化医疗等领域有着应用潜力。并且，项目计划发表的期刊、会议论文与专利，在项目实施过程中均已超额完成。