Tumor-spheroid based migration assay

Tumor-spheroid based migration assay was performed as published with minor changes [51]. Briefly, 1 × 10³ MCF7 cells per well were dispensed into ultra-low attachment 96-well plates (Corning-Ref:7007) and incubated for 5 days. Flat-bottomed, 96-well plates were coated with 0.1% (v/v) gelatin (Sigma) in sterile water for 1 h at 37° C. A total of 200 μl/well of culture medium supplemented with 2% (v/v) FBS was then added. 5 days old spheroids were transferred into the prepared ‘migration’ plate and were allowed to adhere. Images were acquired with a Leica DM-IL microscope. The total area of each spheroid was measured at days 5 and 7 using the ImageJ software.