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Small. 2019 Apr 18:e1901001. doi: 10.1002/smll.201901001. [Epub ahead of print]

Screening Therapeutic Agents Specific to Breast Cancer Stem Cells Using a Microfluidic Single-Cell Clone-Forming Inhibition Assay.

Lin D1,2,3,4, Li P1,5, Feng J1, Lin Z2, Chen X2, Yang N1,2,3,4, Wang L5, Liu D1,2,3,4.

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Department of Laboratory Medicine, Guangzhou First People's Hospital, Guangzhou Medical University, Guangzhou, 510180, China.
Department of Laboratory Medicine, Guangzhou First People's Hospital, School of Medicine, South China University of Technology, Guangzhou, 510180, China.
Guangdong Engineering Technology Research Center of Microfluidic Chip Medical Diagnosis, Guangzhou, 510180, China.
Clinical Molecular Medicine and Molecular Diagnosis Key Laboratory of Guangdong Province, Guangzhou, 510180, China.
Department of Pathology, School of Medicine, Jinan University, Guangzhou, 510632, China.


Screens of cancer stem cells (CSCs)-specific agents present significant challenges to conventional cell assays due to the difficulty in preparing CSCs ready for drug testing. To overcome this limitation, developed is a microfluidic single-cell assay for screening breast cancer stem cell-specific agents. This assay takes advantage of the single-cell clone-forming capability of CSCs, which can be specifically inhibited by CSC-targeting agents. The single-cell assay is performed on a microfluidic chip with an array of 3840 cell-capturing units; the single-cell arrays are easily formed by flowing a cell suspension into the microchip. Achieved is a single cell-capture rate of ≈60% thus allowing more than 2000 single cells to be analyzed in a single test. Over long-term suspension culture, only a minority of cells survive and form tumorspheres. The clone-formation rate of MCF-7, MDA-MB-231, and T47D cells is 1.67%, 5.78%, and 5.24%, respectively. The clone-forming inhibition assay is conducted by exposing the single-cell arrays to a set of anticancer agents. The CSC-targeting agents show complete inhibition of single-cell clone formation while the nontargeting ones show incomplete inhibition effects. The resulting microfluidic single-cell assay with the potential to screen CSC-specific agents with high efficiency provides new tools for individualized tumor therapy.


anticancer agents; cancer stem cells; drug screening; microfluidic chips; single cells


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