CAS Key Laboratory of Standardization and Measurement for Nanotechnology, CAS Key Laboratory for Biomedical Effects of Nanomaterials and Nanosafety, CAS Center for Excellence in Nanoscience, National Center for Nanoscience and Technology of China, Beijing, 100190, China.
School of Nanoscience and Technology, Sino-Danish College, University of Chinese Academy of Sciences, Beijing, 100049, China.
Proteomics. 2021 Feb;21(3-4):e2000060. doi: 10.1002/pmic.202000060. Epub 2021 Jan 12.
Single-cell RNA sequencing on circulating tumor cells (CTCs) proves useful to study mechanisms of tumor heterogeneity, metastasis, and drug resistance. Currently, single-cell RNA sequencing of CTCs usually takes three prerequisite steps: enrichment of CTCs from whole blood, characterization of captured cells by immunostaining and microscopic imaging, and single-cell isolation through micromanipulation. However, multiple pipetting and transferring steps can easily cause the loss of rare CTCs. To address this issue, a novel integrated microfluidic chip for sequential enrichment, isolation, and characterization of CTCs at single-cell level, is developed. And, single CTC lysis is achieved on the same chip. The microfluidic chip includes functions of blood clot filtration, single-cell isolation, identification, and target single-cell lysate collection. By spiking tumor cells into whole blood, it is validated that this microfluidic chip can effectively conduct single-cell CTCs RNA sequencing. The approach lays a solid foundation for the analysis of RNA expression profiling of single-cell CTCs.
循环肿瘤细胞 (CTC) 的单细胞 RNA 测序可用于研究肿瘤异质性、转移和耐药性的机制。目前，CTC 的单细胞 RNA 测序通常需要三个前提步骤：从全血中富集 CTCs、通过免疫染色和显微镜成像对捕获的细胞进行特征描述，以及通过微操作进行单细胞分离。然而，多次移液和转移步骤容易导致稀有 CTC 的丢失。为了解决这个问题，开发了一种新型的集成微流控芯片，用于在单细胞水平上对 CTC 进行顺序富集、分离和特征描述。并且，在同一芯片上实现了单个 CTC 的裂解。微流控芯片包括血凝块过滤、单细胞分离、鉴定和目标单细胞裂解物收集等功能。通过向全血中添加肿瘤细胞进行验证，该微流控芯片可以有效地进行单细胞 CTCs RNA 测序。该方法为单细胞 CTC 的 RNA 表达谱分析奠定了坚实的基础。
