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Single-Cell-Derived Tumor-Sphere Formation and Drug-Resistance Assay Using an Integrated Microfluidics

Cancer stem cells (CSCs) play an important role in tumor progression, metastasis, and drug resistance. In vitro assays using CSC-derived tumorspheres have been used to study the properties of CSCs, and potential anti-CSC agents.

In a recent issue of Analytical Chemistry, Pang et. al. report the development of an integrated microfluidics platform for the formation and analysis of single-cell-derived tumorspheres. By using multi-obstacle architecture-looking (MOAL) microstructural matrices within the device, single glioblastoma cells were captured by their biomechanical properties. These single cells were then channeled into micro-well arrays and cultured into tumorspheres. By testing resistance of the tumorspheres to the anti-cancer drug vincristine, the authors showed that smaller and/or more deformable CSCs were more drug-resistant than larger and/or lesser deformable CSCs. Drug sensitivity was determined by monitoring mitochondrial depolarization using JC-1 dye, and by detecting apoptosis with NucView® 488 Caspase-3 Substrate.

This proof-of-concept study offers a new method for studying CSCs, by using a single microfluidics device for the formation and analysis of single-cell-derived tumorspheres.

 

Dual Apoptosis Assay with NucView® 488 Caspase-3 Substrate & Annexin V
Apoptotic Jurkat cell stained with NucView® 488 (green) and CF®647 Annexin V (magenta).

 

 

 

Full Citation

Pang, L., Ding, J., Ge, Y., Fan, J., & Fan, S. K. (2019). Single-Cell-Derived Tumor-Sphere Formation and Drug-Resistance Assay Using an Integrated Microfluidics. Analytical chemistry.
https://pubs.acs.org/doi/10.1021/acs.analchem.9b01084