Costar 24-well cell migration plates with polycarbonate membranes with 8-µm pore size (Costar Corporation, Cambridge, MA) were used for this assay.
Even higher throughput is allowed by specific migration plates developed by Platypus Technologies (Gough et al. 2011).
For migration assays, lower wells of the SIM plates (migration plates) were filled with growth medium (20% fetal calf serum in RPMI).
For migration experiments, MDA-MB-231 cells were seeded at the same density as for invasion assays onto the upper chamber of 24-well migration plates and 600 μl of serum-free DMEM + 0.1% bovine serum albumin, or serum-free medium + CXCL12 was added to the lower chambers of migration plates and incubated for 24 h.
For migration assays, lower wells of the SIM plates (migration plates) were filled with growth medium (20% FCS in RPMI1640) as a chemoattractant, and cells were plated in the top wells in empty buffer (RPMI1640 only).
Fibronectin-dependent chemotaxis was assessed using transwell migration plates (5.0 μm pores, Corning Life Sciences, Acton, MA, USA) coated with 10 μg/mL of fibronectin (Invitrogen, Carlsbad, CA, USA), a ligand for the β1 integrin [ 25]. 1 × 106 PBMCs were incubated with or without 10 μg/mL of epratuzumab and allowed to migrate for 90 minutes at 37°C and 5% CO2 using transwell migration assays.
The implications of the high degree of migration and plate deformation for long-term survival are not known.
In contrast, both PC-3 and mPanc96 cells failed to show significant differences in migration when plated on soft versus more rigid substrates (Fig. 7C).
Additionally, the rigidity-dependent lines showed a marked decrease in cell spreading and migration when plated on soft versus rigid substrates, and at least one of the cell lines (A549) exhibited a rigidity-dependent regulation of E-cadherin expression and reversible modulation of epithelial and mesenchymal phenotype.
