After 7 days of preconditioning in TCM or CCM, 6.5 × 104 FRCs were seeded onto PCF 0.4 μm pore transwell inserts (Millipore) and cultured overnight in full growth media. To ensure the presence of confluent monolayers, inserts were stained with CellMaskTM plasma membrane stain (Molecular Probes). Inserts were transferred to a fresh 24 well plate and 500 μl basal media was added to the lower chamber. 200 μl media containing 2 % FBS and dextrans at 10,000 MW (Cascade Blue) 70,000 MW (TRITC) and 500,000 MW (FITC; all Life technologies) at a final concentration of 1 mg/ml were added to the upper chamber and fluorescence was measured of the lower chambers’ media after 22h.
Pcf 0.4 μm pore transwell inserts
Manufactured by Merck Group
The PCF 0.4 μm pore transwell inserts are a laboratory equipment product designed for cell culture applications. These inserts feature a polycarbonate membrane with a pore size of 0.4 μm, which allows for the study of cell migration, transport, and other cellular interactions across a semi-permeable barrier.
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Lab products found in correlation
2 protocols using pcf 0.4 μm pore transwell inserts
1
Transwell Permeability Assay for FRCs
2
Transwell Permeability Assay for FRCs
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After 7 days of preconditioning in TCM or CCM, 6.5 × 104 FRCs were seeded onto PCF 0.4 μm pore transwell inserts (Millipore) and cultured overnight in full growth media. To ensure the presence of confluent monolayers, inserts were stained with CellMaskTM plasma membrane stain (Molecular Probes). Inserts were transferred to a fresh 24 well plate and 500 μl basal media was added to the lower chamber. 200 μl media containing 2 % FBS and dextrans at 10,000 MW (Cascade Blue) 70,000 MW (TRITC) and 500,000 MW (FITC; all Life technologies) at a final concentration of 1 mg/ml were added to the upper chamber and fluorescence was measured of the lower chambers’ media after 22h.
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