Cell Aggregate Analysis Protocol

Cells were plated at similar concentrations and grown for 2 days on 35 mm CellBind culture dishes (Corning, Corning, NY, USA) [7 (link)]. At confluency, cells were washed twice and then resuspended in media. Cells were detached with a cell scraper, and then cell aggregates were dissociated by pipetting fifteen times. Images (30–35 fields/dish) were acquired on an Olympus IX 71 microscope with a 10× objective. Particles (≥5 cells/aggregate) were counted, and their areas determined by employment of Image J software. Data is given as the mean ± S.E. and n denotes the number of particles. Statistical comparison was evaluated via student’s t-test. Statistical significance was considered at p < 0.05.