Cell monolayers in individual wells were scraped with a sterile pipette tip (100 μl) after the different groups of ICC cells (1 × 106 cells/well) were grown to 90% confluence. The cells were incubated for 24 h and photographed with a digital camera (Leica, Heerburg, Germany) at 0 h and 24 h after wounding. The distance of cell migration into the denuded area was used to determine the degree of wound healing [24 (link)].
The different groups of ICC cells (1 × 104 cells/well) were seeded in the upper chambers of 24-well Transwell plates containing Matrigel-coated membranes (BD Biosciences, San Jose, CA, USA). Complete medium was added to the bottom chambers. After 36 h of incubation, uninvaded cells were removed from the top surface of the membrane in the upper chamber with a cotton swab. Invaded cells on the underside of the membrane in the upper chamber were fixed with 4% paraformaldehyde, stained with 1% crystal violet and counted under a microscope in a blinded manner [24 (link)].
The different groups of ICC cells (1 × 104 cells/well) were seeded in the upper chambers of 24-well Transwell plates containing Matrigel-coated membranes (BD Biosciences, San Jose, CA, USA). Complete medium was added to the bottom chambers. After 36 h of incubation, uninvaded cells were removed from the top surface of the membrane in the upper chamber with a cotton swab. Invaded cells on the underside of the membrane in the upper chamber were fixed with 4% paraformaldehyde, stained with 1% crystal violet and counted under a microscope in a blinded manner [24 (link)].
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