For the cell migration assay, cells in the logarithmic growth phase were seeded into the upper transwell chamber insert (Corning Inc., Corning, NY, USA) at a density of 2 × 104 cells per well. The chamber was placed in a 24‐well plate, in which the upper chamber contained serum‐free cell culture medium and the lower chamber contained 10% fetal bovine serum (FBS). The culture was continued for 24 h. The medium was discarded, and the cells were stained with a crystal violet solution to observe the number of migrated cells. For the cell invasion assay, Matrigel (Corning Inc.) was uniformly applied to the upper chamber prior to cell inoculation. The other procedures matched those of the cell migration experiment.
Upper transwell chamber insert
Manufactured by Corning
Sourced in United States
The Upper Transwell Chamber Insert is a laboratory equipment component used in cell culture experiments. It serves as a platform for studying cell migration, permeability, and other cellular interactions across a permeable membrane. The insert is designed to fit within a Transwell plate, creating a controlled environment for such analyses.
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8 protocols using upper transwell chamber insert
1
Cell Migration and Invasion Assay
2
Transwell Assay for Cell Migration and Invasion
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Cell migration and invasion ability was measured by Transwell assay. For cell migration assay, cells in logarithmic growth phase were seeded at the upper transwell chamber insert (Corning, USA) at a density of 2 × 104 cells per well. The chamber was placed in a 24‐well plate in which the upper chamber contained serum‐free cell culture medium and the lower chamber contained 10% FBS complete medium. The culture was continued for 24 hours. The medium was discarded, and stained with a crystal violet solution to observe the number of migrated cells. For cell invasion assay, Matrigel gel was uniformly applied to the upper chamber in advance, and then the cells were inoculated. The other procedures were the same as those in the cell migration experiment.
3
Cell Migration Assay Protocol
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Cells in logarithmic growth phase were seeded at the upper transwell chamber insert (Corning, United States) at a density of 2 × 104 cells per well. The chamber was placed in a 24-well plate in which the upper chamber contained serum-free cell culture medium and the lower chamber contained 10% FBS complete medium. The culture was continued for 24 h. The medium was discarded, and stained with a crystal violet solution to observe the number of migrated cells.
4
Transwell Migration Assay Protocol
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A transwell migration assay was conducted as per our previous study (30 (link)). Briefly, cells in the logarithmic growth phase were seeded in the upper transwell chamber insert (Corning, USA) at a density of 2×104 cells per well. To inhibit cell division, serum-free medium containing 1 mM mitomycin was used to replace the complete medium. A 24-well plate was used to place the chamber, where the upper chamber containing FBS-free medium and the lower chamber containing 10% FBS complete medium. Cells cultured for 24 hours. Then discarded the medium and stained the cells with 1% crystal violet solution in order to present the amount of migrated cells.
5
Transwell Migration and Invasion Assay
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A total of 5×104 cells were resuspended in 200 μL of serum-free culture medium and seeded in the upper transwell chamber insert (Corning Inc., Corning, NY, USA) coated (for invasion analysis) or uncoated (for migration analysis) with Matrigel (BD Biosciences). The lower chamber was filled with 600 µL medium containing 10% FBS. After 48 hrs of incubation, the cells remaining on the top membrane were removed, and the cells that passed through the filter were fixed with 4% paraformaldehyde and stained with 0.5% crystal violet. The number of stained cells was counted under a microscope.
6
Cell Migration Assays for Wound Healing
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Wound healing and transwell assays were performed to measure the ability of cell migration. For the wound healing assay, cells were seeded in 6-well plates and cultured for 24 and 48 hours, respectively. A pipette tip (200 μl) was used to make a straight scratch. Cell wound images were taken by a microscope at 0, 24 and 48 hours for examining wound healing.
For the transwell migration assay, 2×104 cells were seeded in the upper transwell chamber insert (Corning, USA). The lower chamber was filled with the complete culture medium containing 10% serum. Cells were allowed to migrate towards the serum gradient for 24 hours. Migrated cells were stained with 1% crystal violet and counted using a phase-contrast microscope. Five random fields were counted per experiment.
For the transwell migration assay, 2×104 cells were seeded in the upper transwell chamber insert (Corning, USA). The lower chamber was filled with the complete culture medium containing 10% serum. Cells were allowed to migrate towards the serum gradient for 24 hours. Migrated cells were stained with 1% crystal violet and counted using a phase-contrast microscope. Five random fields were counted per experiment.
7
Cell Migration Assay Protocol
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Cells in logarithmic growth phase were seeded at the upper transwell chamber insert (Corning, USA) at a density of 2 × 104 cells per well. The chamber was placed in a 24- well plate in which the upper chamber contained serum‐free cell culture medium and the lower chamber contained 10% FBS complete medium. The culture was continued for 24 h. The medium was discarded, and stained with a crystal violet solution to observe the number of migrated cells.
8
Cell Migration Assays for Wound Healing
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Wound healing and transwell assays were performed to measure the ability of cell migration. For the wound healing assay, cells were seeded in 6-well plates and cultured for 48 hours. A pipette tip (200 μl) was used to make a straight scratch. Cell wound images were taken by a microscope at 0 and 48 hours for examining wound healing. For the transwell migration assay, 2 × 104 cells were seeded in the upper transwell chamber insert (Corning, USA). The lower chamber was filled with a complete culture medium containing 10% serum. Cells were allowed to migrate towards the serum gradient for 24 hours. Migrated cells were stained with 1% crystal violet and counted using a phase-contrast microscope. Five random fields were counted per experiment.
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