Dynamic Generation and Culture of 3D Cell Aggregates

3D cell aggregates were generated within the microfluidic device. The following parameters were used for the seeding phase: 10M cells/ml, 2.5 μl/min, 20 min. After seeding, devices were incubated at 37°C, 5% CO₂ and perfused with serum-free chondrogenic medium (Lopa et al., 2013) using an intermittent flow rate (1 min perfusion each hour). Three flow rates were tested: 20, 100, and 500 μl/min. Samples were cultured for 14 days.

Computational models were gleaned on the different aggregate geometries generated upon the tested flow conditions. Specifically, three models were developed: a filled chamber, a disk-shaped aggregate, and a spherical aggregate. The filled chamber model was generated creating a 3D surface above the concave chamber. The other models were developed by designing solid bodies inside the chamber: a disk (Ø_max 1.9 mm, height 1.1 mm) or a solid sphere (Ø 1 mm).