Cell migration and invasion assays68 (link) were performed in vitro using Boyden chamber transwell membranes with a permeable polycarbonate membrane containing 8 μm pores separating the two chambers (Corning). For the invasion assay, the membranes were first coated with 500 μL of Matrigel (Corning) diluted in serum-free medium (1:5) at 37°C for 1 h. One milliliter of migration buffer (medium containing 10% FBS) was added to the lower chamber and incubated for 30 min at 37°C. A total of 1.8 × 104 cells was plated in the upper chamber in 0.5% FBS and incubated for 24 h at 37°C. After 24 h the transwell was removed from the plate, 750 μL of 70% ethanol was added per well in a 24-well plate, and the transwell was placed into the well to fixate the cells which had migrated through the membrane and attached to the transwell. The transwell was removed and dried for 10 min. Next, 750 μL of 0.2% crystal violet (Sigma-Aldrich) was added to the well and incubated for 30 min. Migrated cells were imaged under an inverted microscope and counted using ImageJ.
Permeable polycarbonate membrane
Manufactured by Corning
Sourced in United States
The Permeable polycarbonate membrane is a laboratory equipment product used for filtration and separation processes. It is a thin, porous membrane made of polycarbonate material, allowing the selective passage of certain molecules or particles while retaining others. The membrane's permeability and pore size can be customized to meet the specific requirements of various laboratory applications.
Automatically generated - may contain errors
Lab products found in correlation
Matrigel, by Corning (1 mentions)
Coulter z1, by Beckman Coulter (1 mentions)
Transwell insert, by Corning (1 mentions)
150 cm2 cell culture flask, by Corning (1 mentions)
Fetal bovine serum (fbs), by Merck Group (1 mentions)
Htb 37, by American Type Culture Collection (1 mentions)
Oleic acid, by Avanti Polar Lipids (1 mentions)
4 protocols using permeable polycarbonate membrane
1
Boyden Chamber Migration and Invasion Assay
2
Neutrophil Transmigration Assay
3
Caco-2 Cell Culture for Transport Studies
4
Fluorescent Labeling of Schistosoma Eggs
Check if the same lab product or an alternative is used in the 5 most similar protocols
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HepG2 cells (stock ordered in 2019, CLS # 330198, expanded and stored as cryostocks for consistent quality in culture for up to 10 passages per cryostock) were stimulated with 15 μg/ml SEA and/or 10 mM reduced L-gluthatione (GSH) for the indicated time points. For co-culture assays, HepG2 cells were treated with 400 μM fluorescently labeled oleic acid (Avanti Polar Lipids, Alabaster, AL, USA, #810259C) for 24 h and washed three times with PBS prior to co-culturing with 100 S. mansoni eggs per well (24-well plate) for a further 24 h. In vitro-laid or liver-extracted eggs were co-cultured in the same compartment or in transwells with a permeable polycarbonate membrane (Corning, New York, USA), as indicated in the figure legends. Subsequently, eggs were washed three times and analyzed directly or after fixation with 1% formaldehyde by confocal laser scanning microscopy.
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