Figure 2
The efficiency in seeding cells and generating 3D spheroids between the addition of cells through the seeding port and perfusing cells through the microfluidic device. Transillumination images of HT29 cells in suspension that were (A) added directly through the top cell seeding port and (B) seeded through fluid flow into the microfluidic device. For both conditions, the microfluidic devices were kept under static conditions for 48 h to allow 3D HT29 spheroids to form. This demonstrates the importance of seeding cells directly to the location of the wells rather than through flow. Images are representative of at least 3 independent experiments. (C) Seeding cells directly into the wells is more effective at successfully culturing 3D spheroids in all the wells of the microfluidic device as compared to seeding cells through flow (3 replicates per experiment). (D) 20 μL of HT29 cells (2 × 107cells/mL) were seeded through the top mounted cell seeding port using a pipette. 5 wells from each 5 × 5 array of wells were randomly selected and the number of cells seeded into the wells were manually counted (5 wells/replicates per experiment). Data shown represent means with standard deviation of 3 independent experiments for both spheroid counting and cell counting experiments.
