Cell migration and invasion were assessed with modified Boyden chamber (Becton Dickinson Labware) assays. Briefly, cells, approximately 1.0 × 105, were plated into the upper chamber of a polycarbonate Transwell filter chamber, with or without matrigel (Corning, NY, USA). After 24 h, cells that did not migrate were removed from the top side of the inserts with a cotton swab. Cells that migrated to or invaded the underside of the inserts were fixed in 4% paraformaldehyde, stained with crystal violet, and the migratory cells were counted under a light microscope. Cells were counted in 5 random fields per insert. Three independent experiments were carried out [71 (link)].
Polycarbonate transwell filter chamber
Manufactured by Corning
Sourced in United States
The Polycarbonate Transwell filter chamber is a laboratory equipment used for cell culture applications. It consists of a polycarbonate membrane that acts as a barrier, allowing for the study of cell migration, permeability, and other cellular interactions in a controlled environment.
Automatically generated - may contain errors
6 protocols using polycarbonate transwell filter chamber
1
Transwell Assay for Cell Migration and Invasion
2
Cell Migration Assay Protocol
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A total of 1×105 cells were cultured in the upper chamber of the polycarbonate Transwell filter chamber (Corning) containing Matrigel (Becton, Dickinson and Company, Shanghai, China) and incubated for 16 h. Cells on the lower surface of the chamber were fixed with 4% paraformaldehyde and stained with 0.25% crystal violet (Solarbio Technology Co., Ltd. Beijing, China). Cells on the upper surface were cleaned. Cells were observed and counted in 5 random 100×fields per well using a microscope (Olympus Corporation, Beijing, China).
3
Boyden Chamber Cell Migration Assay
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Cell migration was assessed with modified Boyden chamber (Becton Dickinson, Franklin Lakes, NJ, USA) assays. Briefly, approximately 1.0×105 cells were plated into the upper chamber of a polycarbonate Transwell filter chamber (Corning Inc., Corning, NY, USA). After 24 h, cells that did not migrate were removed from the top side of the inserts with a cotton swab. Migrated cells were stained with crystal violet and fixed in 4% paraformaldehyde followed by counting under a microscope. Cells were counted in five random fields per insert. Three independent experiments were carried out.
4
Cell Invasion Assay with Transwell
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A total of 1 × 105 cells were seeded into the upper chamber of a polycarbonate transwell filter chamber (Corning) and incubated for 22 h. For the invasion assays, the upper chamber was coated with Basement Membrane (R&D, Minneapolis, MN). Cells on the lower membrane surface were fixed in 4% paraformaldehyde, stained with crystal violet and counted (5 random 100× fields per well). Three independent experiments were performed and the data are presented as the average ± SD.
5
NPC Cell Invasion Assay with EVs
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A total of 1 × 105 NPC cells were seeded into the Matrigel (200 μg/ml)‐coated upper chamber of a polycarbonate transwell filter chamber (Corning) and separately co‐incubated with H‐EVs (5 μg/ml) or L‐EVs (5 μg/ml) for 24 h. After non‐invading cells were removed from the top chamber with a cotton swab, cells on the lower membrane surface were fixed in 4% paraformaldehyde, stained with 0.1% crystal violet, and photographed in three independent 20 × fields for each well.
6
Cell Invasion Assay Protocol
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A total of 1 × 105 cells were plated into the upper chamber of a polycarbonate transwell filter chamber (Corning, New York, NY, USA) and incubated for 10 h. For invasion assay, the upper chamber was coated with Matrigel (R&D, Minneapolis, MN, USA) and incubated for 24 h. The non-invading cells were gently removed with a soft cotton swab, and the cells that had invaded to the bottom chamber were fixed, stained, photographed and counted.
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