An airway organoid (AO) model was generated according to our previous report (Sano et al., 2022 (link)). Briefly, normal human bronchial epithelial cells (NHBEs, Cat# CC-2540, Lonza) were used to generate AOs. NHBEs were suspended in 10 mg/ml cold Matrigel growth factor reduced basement membrane matrix (Corning, Cat# 354230). Fifty microliters of cell suspension were solidified on prewarmed cell culture-treated multiple dishes (24-well plates; Thermo Fisher Scientific, Cat# 142475) at 37 °C for 10 min, and then, 500 μl of expansion medium was added to each well. AOs were cultured with AO expansion medium for 10 days. For maturation of the AOs, expanded AOs were cultured with AO differentiation medium for 5 days. In experiments evaluating the antiviral drugs (see “Antiviral drug assay using SARS-CoV-2 clinical isolates and AOs” section below), AOs were dissociated into single cells and then were seeded into a 96-well plate.
Nhbes
Manufactured by Corning
NHBEs (Normal Human Bronchial Epithelial cells) are primary lung cells derived from healthy human donors. They serve as a model system for studying respiratory biology and function.
Automatically generated - may contain errors
3 protocols using nhbes
1
Generation of Airway Organoids for Antiviral Drug Evaluation
2
Generation and Infection of Airway Organoid-ALI Model
Check if the same lab product or an alternative is used in the 5 most similar protocols
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An airway organoid (AO) model was generated according to our previous report2 ,10 (link),37 (link),38 . Briefly, normal human bronchial epithelial cells (NHBEs, Cat# CC-2540, Lonza) were used to generate AOs. NHBEs were suspended in 10 mg/ml cold Matrigel growth factor reduced basement membrane matrix (Corning, Cat# 354230). Fifty microliters of cell suspension were solidified on prewarmed cell culture-treated multiple dishes (24-well plates; Thermo Fisher Scientific, Cat# 142475) at 37 °C for 10 min, and then, 500 μl of expansion medium was added to each well. AOs were cultured with AO expansion medium for 10 days. For maturation of the AOs, expanded AOs were cultured with AO differentiation medium for 5 days.
The AO-ALI model (Fig.5c ) was generated according to our previous report10 (link),91 (link). For generation of AO-ALI, expanding AOs were dissociated into single cells, and then were seeded into Transwell inserts (Corning, Cat# 3413) in a 24-well plate. AO-ALI was cultured with AO differentiation medium for 5 days to promote their maturation. AO-ALI was infected with SARS-CoV-2 from the apical side.
The AO-ALI model (Fig.
3
Generation and Infection of Airway Organoids
Check if the same lab product or an alternative is used in the 5 most similar protocols
+ Expand
An airway organoid (AO) model was generated according to our previous report2 (link),78 (link). Briefly, normal human bronchial epithelial cells (NHBEs, Cat# CC-2540, Lonza) were used to generate AOs. NHBEs were suspended in 10 mg/ml cold Matrigel growth factor reduced basement membrane matrix (Corning, Cat# 354230). Fifty microliters of cell suspension were solidified on prewarmed cell culture-treated multiple dishes (24-well plates; Thermo Fisher Scientific, Cat# 142475) at 37°C for 10 min, and then, 500 μl of expansion medium was added to each well. AOs were cultured with AO expansion medium for 10 days. For maturation of the AOs, expanded AOs were cultured with AO differentiation medium for 5 days.
The AO-ALI model (Fig.5f ) was generated according to our previous report2 (link),78 (link). For generation of AO-ALI, expanding AOs were dissociated into single cells, and then were seeded into Transwell inserts (Corning, Cat# 3413) in a 24-well plate. AO-ALI was cultured with AO differentiation medium for 5 days to promote their maturation. AO-ALI was infected with SARS-CoV-2 from the apical side.
The AO-ALI model (Fig.
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