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Multi-size spheroid formation using microfluidic funnels.

M Marimuthu¹, N Rousset, A St-Georges-Robillard

¹Department of Engineering Physics, Polytechnique Montréal, Canada. thomas.gervais@polymtl.ca.

|December 7, 2017

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This study introduces a microfluidic platform for automated, size-controlled spheroid formation using microfluidic funnels. This technology enables precise cell seeding for 3D biological assays and drug discovery applications.

Area of Science:

Biotechnology
Microfluidics
3D Cell Culture

Background:

Spheroid formation is crucial for 3D biological assays and drug discovery.
Existing methods for spheroid generation often lack precise size control and automation.
Microfluidic platforms offer potential for improved spheroid production but require further development.

Purpose of the Study:

To develop an automated microfluidic platform for generating multi-size spheroids with controlled cell seeding.
To introduce technological innovations for enhanced cell seeding control and independent droplet manipulation.
To enable on-chip formation, treatment, staining, and imaging of spheroids for streamlined analysis.

Main Methods:

A novel microfluidic chip design utilizing a

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microfluidic funnel

layer for precise cell metering.

Single inlet loading with constant cell concentration for reproducible spheroid generation.

Hydrophobic polydimethylsiloxane (PDMS) surface for device stability and long-term storage.

Main Results:

Achieved enrichment factors up to 37× for precise cell seeding.
Successfully generated viable spheroids with diameters ranging from 230-420 μm (OV90) and 280-530 μm (TOV112D).
Demonstrated the platform's capability for on-chip spheroid formation, treatment, and analysis.

Conclusions:

The developed microfluidic platform with microfluidic funnels enables automated, size-controlled spheroid formation.
This technology, termed single inlet multi-size spheroid (SIMSS) chips, is suitable for 3D biological assays requiring size-dependent responses.
Potential applications include chemoresponse assays, photodynamic therapy, and drug transport characterization in drug discovery.