Radius cell migration assay kit

This assay was performed to assess the effect of hGHR antagonist on hGH-induced migration property of the cancer cells. The Radius Cell Migration Assay kit from Cell Biolabs (Cell Biolabs #CBA-125) was used for migration assay, and experiments were performed as per manufacturer’s protocol. Briefly, the assay was performed using 24-well plates containing a nontoxic, 0.68 mm biocompatible hydrogel spot that is present at the center of the well, where cells cannot attach. Prior to plating, cells were treated for 48 h, trypsinized, counted, and seeded at 5000 cells/well in the pretreated hydrogel spot containing 24-well plate. The hydrogel spot was gently removed after 24 h incubation at 37 °C/5% CO2, and cells were allowed to migrate for up to 48 h while images were captured every 24 h using a 4× objective (total magnification 40×) employing an inverted Olympus IX70 microscope fitted with a Retiga 1300 camera (QImaging). The total uncovered area at the beginning and end of assay were quantitated using ImageJ software. Experiments were done in triplicates.

The migration ability of HCT116 cells was measured with a gap closure assay using a Radius™ cell migration assay kit (CBA-127, Cell Biolabs). Equal number of cells (10,000 per well) were seeded in 10 wells each group in a 384-well plate. When cells were confluent, the RADIUS gel spot was removed according to the manufacturer's protocol in order to simulate a wound and the cells were cultured for 2 days. The pre-migration and daily post-migration images were captured using an invert fluorescent microscope DMI6000B (Leica). Areas were measured using the “outline” function on the Image J program. The percentage of gap areas over time was calculated by dividing the original RADIUS open.

The Radius™-Cell-Migration Assay Kit (Cell Biolabs, San Diego, CA, USA) utilizes a 24-well plate to monitor the two-dimensional migratory properties of MPCs. Each well contains a 0.68 mm biocompatible hydrogel spot where cells cannot attach. For this experiment, 5 × 10⁴ MPCs were seeded into the Radius™-Cell-Migration well and attached outside of the gel-coated area. After 24 h of culturing, the hydrogel was removed to expose a cell-free region to study cell migration with subsequent closure of the cell-free space. The cells were stimulated either by 10 µg/mL rhTFF3 or 20 µl/mL platelet-rich plasma concentrate (PRP) in DMEM supplemented with 10% FCS and 1% penicillin-streptomycin to yield a total volume of 1 ml per well. PRP was prepared as described previously [35 (link)]. The ingrowth of cells was compared to negative controls at defined time points. Microscopic images were performed in bright field (Axiophot (HBO100), Zeiss, Jena, Germany) at 0 h, 8 h, 24 h, and 48 h (not shown) to document cell migration.