A Microfluidic Chip for Efficient Circulating Tumor Cells Enrichment, Screening, and Single-Cell RNA Sequencing.

Abstract

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.

• Authors: Fanghao Shi, Fei Jia, Zewen Wei, Yan Ma, Zhiguo Fang, Weikai Zhang, Zhiyuan Hu
• Journal: Proteomics (Proteomics) Vol. 21 Issue 3-4 Pg. e2000060 (02 2021) ISSN: 1615-9861 [Electronic] Germany
• PMID: 33219587 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: © 2020 The Authors. Proteomics published by Wiley-VCH GmbH.

Topics

• Cell Line, Tumor
• Cell Separation
• Humans
• Microfluidic Analytical Techniques
• Microfluidics
• Neoplastic Cells, Circulating
• Sequence Analysis, RNA