The DLD-1 and SW48 cells were transfected using siYY1 and siControl 24 h before the gap closure assay. Cells of appropriate density (2×104 cells/well for DLD-1 and 15×104 cells/well for SW48) and 100% confluence in the monolayer were seeded into each well of a culture insert (cat. no. 81176; Culture-Insert 2 Well in µ-Dish; Ibidi GmbH). After 24 h of incubation at 37°C, the culture insert was removed and the dish was filled with complete medium. Images of the cell-free gaps were captured using an inverted light microscope (Axio Observer Z1; Carl Zeiss AG). The images were captured in three fields per well at each point in time (DLD-1, 24 h; and SW48, 96 h after removing the culture-insert). The cell-free gaps were measured using ImageJ software version 1.53k (National Institutes of Health) and the percentage of cell-free gaps was compared with that at 0 h.
Culture insert 2 well in dish
Manufactured by Ibidi
Sourced in Germany
The Culture-Insert 2 Well in µ-Dish is a type of cell culture dish designed by Ibidi. It features two individual culture wells within a single dish format. The core function of this product is to provide a controlled environment for the growth and observation of cells.
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Lab products found in correlation
4 protocols using culture insert 2 well in dish
1
Wound Healing Assay with siYY1 Transfection
2
Cell Migration Assay with HT-29 and LS174T
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Wound-healing assays were used to measure cell migration. The single-cell clones of HT-29 and transiently transfected LS174T cells were seeded in the Culture-Insert 2 Well in µ-Dish (ibidi GmbH, Martinsried, Germany, #81176) at a density of 3 × 104 cells per well. Then, the culture-insert was removed, and the media were exchanged with fresh serum-reduced media (2% FBS). The plate was photographed at different time points, and the insert space was quantified by imageJ, v1.53.
3
Wound Closure Assay with NaB
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Cells were allowed to adhere to the well and grow to greater than 90% confluence in Culture-Insert 2 Well in µ-Dish (80206, ibidi GmbH, Gräfelfing, Germany). Then, cells were treated with 2 mM NaB, the wall between cells was removed, and cultured for 12 h. The changes in the wound area were monitored under a CKX53 phase contrast microscope (Olympus, Tokyo, Japan). The migration distance of each group was analyzed using ImageJ software comparing the area observed at 0 h.
4
MCF7 Cell Migration Assay
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Cell migration assays were performed using the Culture-Insert 2 well in µ-Dish (ibidi GmbH, Martinsried, Germany).
Briefly, 25,000 MCF7 cells were plated in each well and cultured for 24 h. The day after, the culture insert was removed and the cells were washed with PBS before treating them with respective DGKα inhibitors (10 µM) or DMSO for 15 h in complete medium (DMEM 10% FBS + 1% penicillin/streptomycin), while medium without FBS was used as a negative control for migration.
Phase-contrast pictures were taken immediately after treatment (0 h) and after 15 h under 5× magnification.
Finally, wound areas were determined using ImageJ software (NIH, , Bethesda, MD). Wound reduction was calculated by using the following formula: (wound area at 15 h/wound at 0 h)×100, the values obtained were expressed as the percentage of wound area compared to the initial area.
Briefly, 25,000 MCF7 cells were plated in each well and cultured for 24 h. The day after, the culture insert was removed and the cells were washed with PBS before treating them with respective DGKα inhibitors (10 µM) or DMSO for 15 h in complete medium (DMEM 10% FBS + 1% penicillin/streptomycin), while medium without FBS was used as a negative control for migration.
Phase-contrast pictures were taken immediately after treatment (0 h) and after 15 h under 5× magnification.
Finally, wound areas were determined using ImageJ software (NIH, , Bethesda, MD). Wound reduction was calculated by using the following formula: (wound area at 15 h/wound at 0 h)×100, the values obtained were expressed as the percentage of wound area compared to the initial area.
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