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Issue 19, 2016
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Scaling and automation of a high-throughput single-cell-derived tumor sphere assay chip

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Abstract

Recent research suggests that cancer stem-like cells (CSCs) are the key subpopulation for tumor relapse and metastasis. Due to cancer plasticity in surface antigen and enzymatic activity markers, functional tumorsphere assays are promising alternatives for CSC identification. To reliably quantify rare CSCs (1–5%), thousands of single-cell suspension cultures are required. While microfluidics is a powerful tool in handling single cells, previous works provide limited throughput and lack automatic data analysis capability required for high-throughput studies. In this study, we present the scaling and automation of high-throughput single-cell-derived tumor sphere assay chips, facilitating the tracking of up to ∼10 000 cells on a chip with ∼76.5% capture rate. The presented cell capture scheme guarantees sampling a representative population from the bulk cells. To analyze thousands of single-cells with a variety of fluorescent intensities, a highly adaptable analysis program was developed for cell/sphere counting and size measurement. Using a Pluronic® F108 (poly(ethylene glycol)-block-poly(propylene glycol)-block-poly(ethylene glycol)) coating on polydimethylsiloxane (PDMS), a suspension culture environment was created to test a controversial hypothesis: whether larger or smaller cells are more stem-like defined by the capability to form single-cell-derived spheres. Different cell lines showed different correlations between sphere formation rate and initial cell size, suggesting heterogeneity in pathway regulation among breast cancer cell lines. More interestingly, by monitoring hundreds of spheres, we identified heterogeneity in sphere growth dynamics, indicating the cellular heterogeneity even within CSCs. These preliminary results highlight the power of unprecedented high-throughput and automation in CSC studies.

Graphical abstract: Scaling and automation of a high-throughput single-cell-derived tumor sphere assay chip

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Supplementary files

Article information


Submitted
18 Jun 2016
Accepted
26 Jul 2016
First published
04 Aug 2016

Lab Chip, 2016,16, 3708-3717
Article type
Paper

Scaling and automation of a high-throughput single-cell-derived tumor sphere assay chip

Y. Cheng, Y. Chen, R. Brien and E. Yoon, Lab Chip, 2016, 16, 3708
DOI: 10.1039/C6LC00778C

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