Corning® Transwell® polycarbonate membrane cell culture inserts

Protocol tips

Upstream tips	Protocol tips	Downstream tips
- Seed 5 × 10^5 cells/transwell.	- Reconstitute Matrigel basement membrane with 5 μg/cm^2 . - Supplement with 1% FBS medium on both sides of the wells and stiluli on the lower side.

Upstream tips
- Seed 5 × 10^5 cells/transwell.
Protocol tips
- Reconstitute Matrigel basement membrane with 5 μg/cm^2 . - Supplement with 1% FBS medium on both sides of the wells and stiluli on the lower side.

Publication protocol

Cell migration and invasion assays were performed in Transwell™ chambers (6.5 mm diameter polycarbonate filter with 8.0 μm pore size, Corning, NY, USA). For migration assay, cells (A549: 5 × 103/transwell; T47D: 105) were directly seeded on the upper side of the filters. For invasion assay, filters were coated with reconstituted Matrigel basement membrane (Collaborative Research, Waltham, MA, USA) at the following concentrations: A549 cells (105/transwell) with 12.5 μg/cm2; H322 cells (105/transwell) with 5 μg/cm2; SW‐48 cells (105/transwell) with 5 μg/cm2 and T47D cells (105/transwell) with 10 μg/cm2. Culture medium supplemented with 1% FBS was added to both chambers, and stimuli to the lower chamber. After 24 h cells on the filter upper side were mechanically removed, and those migrated onto the lower side were fixed, stained, and photographed with a 10× objective as above. Morphometric analysis was performed using NIH ImageJ software (National Institutes of Health, Bethesda, MD). Data shown are the mean of at least three independent experiments in duplicate ± S.E.M.

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Manufacturer protocol

Download the product protocol from Sigma-Aldrich for Corning® Transwell® polycarbonate membrane cell culture inserts below.

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