The CCHMC CF Research Development Program Translational and Model Systems Cores maintained, and provided wild-type CFTR (wtCFBE41o-) cells as previously described (30 (link)). wtCFBE41o- cells stably transduced with wild-type CFTR were initially generated and described by Gruenert and colleagues, and were initially a kind gift from the University of Alabama at Birmingham (31 (link)). HBE from ten normal healthy donors were obtained from University of North Carolina Airway Cell Core using previously described methods (32 (link)). HBE at passages 2-3 were differentiated and grown at ALI on Corning® 3470 Transwell® polyester membrane cell culture inserts, 6.5 mm diameter, 0.4μm pore size, by the CCHMC Pulmonary Core as described (30 (link)). The CCHMC, Pulmonary Biorepository Core, and University of Cincinnati IRBs approved primary HBE use. Epithelial cells were incubated with NETs from unrelated human donors, at doses of ~5µg/ml, 18h, 37°C, 5% CO2. Higher dose NETs or other incubation periods were used in experiments where indicated. Dornase Alpha (DA), obtained from the CCHMC research pharmacy, was used at a final concentration of 0.5µg/ml to digest the NET DNA structure.
3470 transwell polyester membrane cell culture inserts
Manufactured by Corning
The Corning® 3470 Transwell® polyester membrane cell culture inserts are a laboratory equipment product designed for cell culture applications. The inserts feature a polyester membrane that allows for the exchange of nutrients, gases, and other substances between the upper and lower chambers of the culture system.
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Lab products found in correlation
2 protocols using 3470 transwell polyester membrane cell culture inserts
1
Exposure of Primary HBE to NETs
2
Healthy Human Airway Epithelial Cell Culture and NET Exposure
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The CCHMC CF Research Development Program Translational and Model Systems Cores maintained, and provided wild-type CFTR (wtCFBE41o-) cells as previously described (28) (link). wtCFBE41o-cells stably transduced with wild-type CFTR were initially generated and described by Gruenert and colleagues, and were initially a kind gift from the University of Alabama at Birmingham (27) . HBE from ten normal healthy donors were obtained from University of North Carolina Airway Cell Core using previously described methods (78) . HBE at passages 2-3 were differentiated and grown at ALI on Corning® 3470 Transwell® polyester membrane cell culture inserts, 6.5 mm diameter, 0.4μm pore size, by the CCHMC Pulmonary Core as described (28) (link). The CCHMC, Pulmonary Biorepository Core, and University of Cincinnati IRBs approved primary HBE use. Epithelial cells were incubated with NETs from unrelated human donors, at doses of ~5µg/ml, 18h, 37°C, 5% CO2.
dose NETs or other incubation periods were used in experiments where indicated. Dornase Alpha (DA), obtained from the CCHMC research pharmacy, was used at a final concentration of 0.5µg/ml to digest the NET DNA structure.
dose NETs or other incubation periods were used in experiments where indicated. Dornase Alpha (DA), obtained from the CCHMC research pharmacy, was used at a final concentration of 0.5µg/ml to digest the NET DNA structure.
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