BEBMTM Bronchial Epithelial Cell Growth Basal Medium

Mammalian cell culture media HBEpC

Experiment
Mammalian cell culture media HBEpC
Product
BEBMTM Bronchial Epithelial Cell Growth Basal Medium from Lonza
Manufacturer
Lonza

Protocol tips

Protocol tips
incubated at 37°C, 5% CO2.

Publication protocol

Cell expansion
Three different initial proliferation culture media were used to revive NHBE (LONZA) cells from cryopreservation, seeded as passage one (P1) into T25 cell culture tissue flasks: BEGM growth media (LONZA, Walkersville MD), PneumaCult-Ex (PnC-Ex) media (StemCell, Tukwila WA) or PneumaCult-Ex Plus (PnC-Ex-PLUS) media (StemCell, Tukwila WA). Cells were seeded at ~100,000 cells/T25 flask, and incubated at 37°C, 5% CO2. Once cells reached 70–80% confluency, they were dissociated using TryplE Express dissociation media (ThermoFisher, MA USA). From passage two (P2), different growth conditions were tested, including the use of a Rho kinase inhibitor (Y-27632; R&D Systems, Minneapolis MN) and/or feeder cells (mouse 3T3 fibroblast cells treated with mitomycyin C)

Cells cultured at Air-Liquid Interface (ALI)
Once cells had reached 70–80% confluency, they were dissociated and seeded on 6.5 mm Transwells (Corning; Fisher Scientific) coated with 0.3 mg/mL Collagen type IV from human placenta (Sigma-Aldrich, St. Louis MO). NHBE cells grown in BEGM + ROCK, PnC-Ex or PnC-Ex-PLUS media were seeded at 50,000 cells/well. Respective growth media was used to feed cells until 4–5 days or 100% confluency was reached. Upon reaching confluency, the apical media was removed and the basal media replaced with PneumaCult-ALI medium for all cell conditions. Medium was changed every second day and apical surfaces washed with HBSS (Ca2+, Mg2+) twice per week. All cells were grown until 4 weeks airlifted (37°C, 5% CO2) before various physiological and characterization tests were performed. Epithelial layers were grown from passaged cells until they failed to expand any further.





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Manufacturer protocol

Download the product protocol from Lonza for BEBMTM Bronchial Epithelial Cell Growth Basal Medium below.

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