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A Microfluidic Chip for Efficient Circulating Tumor Cells Enrichment, Screening, and Single‐Cell RNA Sequencing
Author(s) -
Shi Fanghao,
Jia Fei,
Wei Zewen,
Ma Yan,
Fang Zhiguo,
Zhang Weikai,
Hu Zhiyuan
Publication year - 2021
Publication title -
proteomics
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.26
H-Index - 167
eISSN - 1615-9861
pISSN - 1615-9853
DOI - 10.1002/pmic.202000060
Subject(s) - circulating tumor cell , single cell analysis , microfluidics , rna , cell , lysis , microfluidic chip , single cell sequencing , computational biology , immunostaining , biology , microbiology and biotechnology , metastasis , cancer , phenotype , nanotechnology , gene , immunology , exome sequencing , materials science , genetics , immunohistochemistry
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.