恶性肿瘤单细胞电学特性高通量检测方法研究

Tumor heterogeneity, which describes that different tumor cells show distinct metastatic, proliferation and drug resistance, introduces significant challenges in cancer treatment and prognosis. Research into understanding and characterizing heterogeneity can allow for a better understanding of the causes and progression of disease. In the study of tumor heterogeneity, single cell analysis of tumor cells, which focus on studying the single cells’ biophysical and biochemical characteristics is very meaningful. Single cells’ electrical properties (including specific membrane capacitance (capacitance per unit area) and cytoplasm conductivity), which are important single cells’ biophysical properties, may serve as an important dimension to observe cell heterogeneity and have great research value for understanding the tumor heterogeneous. Facing the huge cell number in the human tumor samples, the throughput of the existing methods is still limited, which causes the single cells’ electrical properties of the mainstream tumor cells in the tumor tissue cannot be obtained. Address this issue, this project proposed a new innovative detection method on double channel based on the structure of microtubules and compression, the method is put forward: the core structure of high-throughput accurate detection cell electrical properties -- on double microtubules and compression channel structure, combining the theory of modeling structure equivalent electrical model, can achieve high throughput on malignant tumor cells high precision detection coverage. By exploring the relationship between the electrical properties of single cells and the heterogeneity of tumor cells, this method is expected to provide a new dimension to observe the heterogeneity of tumor cells, and provide a theoretical and theoretical support for further understanding of tumor heterogeneity.

肿瘤异质性即肿瘤内不同细胞的增殖、转移能力和耐药性等方面存在差异，导致肿瘤诊疗和预后面临挑战。针对肿瘤异质性，利用单细胞分析方法，研究肿瘤单细胞生物物理和生物化学特性，可以推动恶性肿瘤的诊疗方案的优化。单细胞电学特性包括细胞膜比电容和细胞质电导率，作为重要的单细胞固有生物物理特性，可能可以作为一个观察细胞异质性的重要维度，具有很大的研究价值。但是，现有方法的检测通量低，不能得到肿瘤组织中大量特异性活体细胞的电学特性，制约了研究进程。本项目创新的提出一种“高通量电学流式细胞术”的检测方法，该方法中提出实现高通量精准检测细胞电学特性的核心结构——双微孔电极和压缩通道结构，和检测结构中单细胞等效电学模型，可以实现对恶性肿瘤组织中活体细胞高覆盖率高通量精准检测。通过探索单细胞电学特性与肿瘤细胞异质性的关系，该方法有望提供一个观察肿瘤细胞异质性的新维度，为进一步全面理解肿瘤异质性提供方法和理论支持。

在基金支持下，本项目研究了一种基于交叉压缩通道敏感原理的单细胞电学特性高通量流式检测方法，将单细胞电学特性的检测通量从每小时数百个细胞提升到10万个细胞，完成了项目设计的目标。进一步，研制成功一台样品进结果出的单细胞电学流式细胞分析仪一台，具有实时检测和计算结果的能力。应用研究发现，在肿瘤研究方面，基于电学特性可以区分不同种类的肿瘤细胞，区分不同转移状态的肿瘤细胞亚型，区分肺癌病人胸水中不同形貌肿瘤细胞；在脑卒中研究方面，应用表明作为一种快速检测的免标记特征，电学特性可被用于区分表征缺血和对照组的大鼠模型中性粒细胞；能够作为一个稳定有效的特征区分不同极化状态的巨噬细胞，并能对药物作用效果进行评价，评价结果与动物实验相一致。.相关成果作为部分工作获得2020年中国仪器仪表学会技术发明二等奖。项目执行中，发表SCI文章4篇，申请专利3项，培养博士生2名，硕士生1名。