Fig. 2.

Migration of ESC- and iPSC-derived fibroblasts towards PDGF stimulation is dependent on PDGFRβ activity and fibronectin. Cells were seeded on a polycarbonate membrane with 8 μM pores, and allowed to migrate for 6 hours towards either no stimulus or 50 ng/μl PDGF-BB in the lower chamber. (A) Migration of BJ, EDK and iPDK cells was determined by washing cells from the upper chamber, fixing cells in 4% paraformaldehyde, and staining cells that had migrated through the membrane with 0.1% Crystal Violet. (B) We quantified these results by counting and averaging migrated cells in three separate experiments and showed a significant increase in migration towards PDGF-BB, and this effect was abrogated with a PDGFRβ blocking antibody. (C) Fibronectin-coated membranes showed the highest level of migratory induction in response to PDGF-BB compared to uncoated or collagen-coated membranes. (D) EDK and iPDK cells with stable knockdown of PDGFRβ showed reduced migration compared to scrambled controls. Significant changes over control conditions were determined using a two-tailed t-test. *P≤0.05.