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Hbtec air liquid interface differentiation medium

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The HBTEC Air-Liquid Interface Differentiation Medium is a specialized cell culture medium designed to facilitate the differentiation of human bronchial epithelial cells (HBECs) at an air-liquid interface. This medium supports the growth and differentiation of HBECs, enabling the cells to develop a polarized, mucociliary phenotype that mimics the in vivo airway epithelium.

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Lab products found in correlation

Stx2 electrode, by World Precision Instruments (3 mentions) Evom voltohmmeter, by World Precision Instruments (3 mentions) Bronchial epithelial growth medium (begm), by Lonza (1 mentions) 6.5 mm transwell, by Corning (1 mentions) Cc 2541, by Lonza (1 mentions)

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HBTECs (5 citations)

4 protocols using hbtec air liquid interface differentiation medium

1

Airway Epithelial Cell Culture Differentiation

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Primary NHBE cells were purchased from Lonza (catalog no. CC-2541; Basel, Switzerland). NHBE cells in a submerged condition were cultured on 6.5-mm Transwell (Corning, NT, USA) using commercially available bronchial epithelial growth medium (BEGM; Lonza) and incubated at 37 °C in a humidified atmosphere with 5% carbon dioxide. When the NHBE cells reached full confluency in an immersed culture condition, the cells were transferred to an ALI culture condition using ALI culture medium (HBTEC Air–Liquid Interface Differentiation Medium; Lifeline Cell Technology, Frederick, MD, USA), as previously described with slight modifications [3 (link), 4 (link), 23 (link), 24 (link)]. The cells were then cultured for 4 weeks in ALI conditions to facilitate polarization and ciliary differentiation for subsequent experiments [25 (link), 26 (link)].
Katsumata M., Fujisawa T., Kamiya Y., Tanaka Y., Kamiya C., Inoue Y., Hozumi H., Karayama M., Suzuki Y., Furuhashi K., Enomoto N., Nakamura Y., Inui N., Maekawa M., Setou M., Watanabe H., Ikegami K, & Suda T. (2022). Effects of long-acting muscarinic antagonists on promoting ciliary function in airway epithelium. BMC Pulmonary Medicine, 22, 186.
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2

Modeling Virus Infection in 3D HBTEC Cultures

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3.3×104 HBTECs were seeded onto 6.5 mm Costar Transwell Cell Culture Inserts (pore size 0.4 μm) and grown submerged in BronchiaLife cell culture medium to 100% confluency. HBTEC Air-Liquid Interface Differentiation Medium (LifeLine Cell Technology) was then added to the basolateral chamber, and media removed from the apical chamber. Cells were grown at ALI for 21 days, emerging 3D cultures washed every 48 hours to remove excess mucus, and transepithelial/transendothelial resistance (TEER) measured using an EVOM volt/ohm meter coupled with an STX2 electrode (World Precision Instruments). Cultures with resistance ≥700 Ω/cm2 were used for experimentation. For infection studies, 3D-ALI-HBTECs were apically inoculated with recHPIV3-JS-NanoLuc, HPIV-1–5F6, HPIV3–9R4, or HPIV3–10L3, (5,000 TCID50 units/well) for 2 hours and washed thrice with media. Treatment with GHP-88309 or volume-equivalent DMSO was administered from the basal chamber. Released virus was collected from the apical chamber every 24 hours. Treatment efficacy was evaluated by TCID50 (recHPIV3-JS-NanoLuc) or TCID50-HA (HPIV-1 and HPIV3 clinical isolates) titration of shed virus.
Cox R.M., Sourimant J., Toots M., Yoon J.J., Ikegame S., Govindarajan M., Watkinson R.E., Thibault P., Makhsous N., Lin M.J., Marengo J.R., Sticher Z., Kolykhalov A.A., Natchus M.G., Greninger A.L., Lee B, & Plemper R.K. (2020). Orally Efficacious Broad-Spectrum Allosteric Inhibitor of Paramyxovirus Polymerase. Nature microbiology, 5(10), 1232-1246.
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3

Differentiated 3D HBTEC Airway Model

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3.3 × 104 HBTECs at a passage number ≤3 were seeded onto 6.5 mm Costar Transwell Cell Culture Inserts (pore size 0.4 μm) and grown submerged in BronchiaLife cell culture medium to 100% confluency. Upon confluency, media was removed from the apical chamber and the media in the basal chamber changed to HBTEC Air-Liquid Interface Differentiation Medium (LifeLine Cell Technology). The cells were grown at air-liquid interface (ALI) for at least 21 days to ensure complete differentiation; media in the basal chamber was changed every 2 days. The developing 3D cultures were washed apically weekly to remove excess mucus. Transepithelial/transendothelial resistance (TEER) was measured using an EVOM volt/ohm meter coupled with an STX2 electrode (World Precision Instruments). Cultures with resistance ≥700 Ω/cm2 were used for experimentation.
Toots M., Yoon J.J., Cox R.M., Hart M., Sticher Z.M., Makhsous N., Plesker R., Barrena A.H., Reddy P.G., Mitchell D.G., Shean R.C., Bluemling G.R., Kolykhalov A.A., Greninger A.L., Natchus M.G., Painter G.R, & Plemper R.K. (2019). Characterization of orally efficacious influenza drug with high resistance barrier in ferrets and human airway epithelia. Science translational medicine, 11(515), eaax5866.
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4

Modeling Virus Infection in 3D HBTEC Cultures

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3.3×104 HBTECs were seeded onto 6.5 mm Costar Transwell Cell Culture Inserts (pore size 0.4 μm) and grown submerged in BronchiaLife cell culture medium to 100% confluency. HBTEC Air-Liquid Interface Differentiation Medium (LifeLine Cell Technology) was then added to the basolateral chamber, and media removed from the apical chamber. Cells were grown at ALI for 21 days, emerging 3D cultures washed every 48 hours to remove excess mucus, and transepithelial/transendothelial resistance (TEER) measured using an EVOM volt/ohm meter coupled with an STX2 electrode (World Precision Instruments). Cultures with resistance ≥700 Ω/cm2 were used for experimentation. For infection studies, 3D-ALI-HBTECs were apically inoculated with recHPIV3-JS-NanoLuc, HPIV-1–5F6, HPIV3–9R4, or HPIV3–10L3, (5,000 TCID50 units/well) for 2 hours and washed thrice with media. Treatment with GHP-88309 or volume-equivalent DMSO was administered from the basal chamber. Released virus was collected from the apical chamber every 24 hours. Treatment efficacy was evaluated by TCID50 (recHPIV3-JS-NanoLuc) or TCID50-HA (HPIV-1 and HPIV3 clinical isolates) titration of shed virus.
Cox R.M., Sourimant J., Toots M., Yoon J.J., Ikegame S., Govindarajan M., Watkinson R.E., Thibault P., Makhsous N., Lin M.J., Marengo J.R., Sticher Z., Kolykhalov A.A., Natchus M.G., Greninger A.L., Lee B, & Plemper R.K. (2020). Orally Efficacious Broad-Spectrum Allosteric Inhibitor of Paramyxovirus Polymerase. Nature microbiology, 5(10), 1232-1246.
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