Spheroids grow with the flow in this microfluidic device

Microfluidic platforms allow scientists to culture 3D cell aggregates — spheroids — to test the efficacy of new drugs and study disease mechanisms in vitro. However, traditional microfluidic platforms have permanently sealed microchannels with fixed geometries that limit researchers’ ability to quickly test different channel configurations and remove the spheroids once grown for downstream analysis. Aljayyousi et al. developed a rapidly reconfigurable microfluidic platform that can be easily changed to accommodate different spheroid growth conditions and can be disassembled for spheroid imaging.

The platform combines the advantages of open well structures for easy cell loading and imaging with those of enclosed microchannels for continuous flow of cell growth media.

“Unlike conventional, permanently sealed microfluidic systems using plasma bonding, our design utilizes a double-sided adhesive layer for the reversible sealing of microwells,” author Yong-Ak Song said. “The adhesive layer between the bottom well layer and top cover layer contains freely customizable channel configurations generated by a laser cutter.”

The authors tested three types of cell lines on their device: mouse embryonic fibroblasts, human-induced pluripotent stem cells, and breast cancer cells. These cells were loaded into open microwells through pipetting, and subsequently, the microfluidic device was sealed with the adhesive layer for continuous perfusion. The team then cultured the spheroids in the device for up to 14 days and performed noninvasive optical coherence tomography (OCT) for viability assessments.

“Spheroid growth, morphology, and viability were assessed through microscopy and OCT imaging,” Song said. “A computational pipeline ensured the robust and accurate measurement of viability from OCT images, validated by standard assays.”

Future studies will optimize well geometries for automatic high-throughput OCT imaging of spheroids.

Source: “A modular and reconfigurable microfluidic device for culturing spheroids under continuous perfusion,” by Hiba Aljayyousi, Sarah Sahloul, Ajymurat Orozaliev, Navajit Baban, Duc-Anh Van, Amani Al Nuairi, Pauline John, Azhar Zam, Piergiorgio Percipalle, and Yong-Ak Song, APL Bioengineering (2025). The article can be accessed at https://doi.org/10.1063/5.0262536 .