Polycarbonate membrane filter inserts with 8 μm pores

Cell migration assays were carried out with transwell filters as described by Liu et al [2 (link)]. Both the Hela cells with Nanog disruption and those without were digested and re-suspended with DMEM medium to 1×10⁵ cells/ml. 200μl cell suspension were seeded into an upper chamber of the polycarbonate membrane filter inserts with 8-μm pores (Corning, USA), which were pre-coated with matrigel. The lower chamber was filled with 500 μl of DMEM medium with 10% FBS. The cells (not migrated) in the upper chamber surface were removed with cotton swabs after incubated at 37°C for 24 and 48 h, and the cells (migrated) on the bottom side of the membrane were fixed with 95% ethanol for 30 min, stained with the 0.1% crystal violet and counted under a microscope. The experiment was performed in triplicates.

Cell migration assays were carried out with transwell filters. U251 or U87-MG cells were digested and re-suspended with DMEM medium to 1 × 10⁵cells/ml. Two hundred microliters cell suspension were seeded into an upper chamber of polycarbonate membrane filter inserts with 8-μm pores (Corning, USA). The lower chamber was filled with 500 μl of DMEM medium with 10% FBS. The cells (not migrated) in the upper chamber surface were removed with cotton swabs after incubated at 37 °C for 24 h, and the cells (migrated) on the bottom side of the membrane were fixed with 95% ethanol for 30 min, stained with the 0.1% crystal violet and counted under a microscope. The experiment was performed in triplicates.