hTert-immortalized human normal airway tracheobronchial epithelial cells, BCi.NS1.1 (58 (link)), were maintained in ExPlus growth media (StemCell). To generate HAEC, BCi.NS1.1 cells were plated (7.5E4 cells/well) on rat-tail collagen type 1-coated permeable transwell membrane supports (6.5 mm; Corning Inc.), immersed in ExPlus growth media in both the apical and the basal chambers. Upon reaching confluence, the medium in the apical chamber was removed (airlift), and the medium in the basal chamber was changed to Pneumacult ALI maintenance media (StemCell). Pneumacult ALI maintenance medium was changed every 2 days for approximately 6 weeks to form differentiated, polarized cultures that resemble in vivo pseudostratified mucociliary epithelium.
Rat tail collagen type 1 coated permeable transwell membrane supports
Manufactured by Corning
Rat-tail collagen type 1-coated permeable transwell membrane supports are a laboratory product designed to facilitate the study of cell migration, invasion, and permeability. The core function of these supports is to provide a controlled, permeable barrier that allows the passage of cells, fluids, and molecules while maintaining a defined microenvironment.
Automatically generated - may contain errors
Lab products found in correlation
Explus growth media, by STEMCELL (2 mentions)
Explus growth media, by Corning (2 mentions)
Pneumacult ali maintenance media, by STEMCELL (2 mentions)
Ficoll paque plus, by GE Healthcare (1 mentions)
Roswell park memorial institute (rpmi), by Thermo Fisher Scientific (1 mentions)
Begm bullet kit, by Lonza (1 mentions)
3 protocols using rat tail collagen type 1 coated permeable transwell membrane supports
1
Differentiation of Airway Epithelial Cells
2
Differentiation of Immortalized Human Airway Epithelial Cells
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hTert-immortalized human normal airway tracheobronchial epithelial cells, BCi.NS1.1 (55 (link)) were maintained in ExPlus growth media (StemCell). To generate HAEC, BCi.NS1.1 were plated (7.5E4 cells/well) on rat-tail collagen type 1-coated permeable transwell membrane supports (6.5mm; Corning Inc), immersed in ExPlus growth media in both the apical and basal chamber. Upon reaching confluency, media in the apical chamber was removed (airlift), and media in the basal chamber was changed to Pneumacult ALI maintenance media (StemCell). Pneumacult ALI maintenance media was changed every two days for approximately 6 weeks to form differentiated, polarized cultures that resemble in vivo pseudostratified mucociliary epithelium.
3
Isolation and Culture of Primary Human Airway Epithelial Cells
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PBMC were isolated from blood using a Ficoll-Paque PLUS (GE Amersham) gradient [66 (link)] and maintained in RPMI (Invitrogen). PBMC were used for functional studies.
Primary human normal airway tracheobronchial epithelial cells from de-identified donors (NHBE) (sex: male and female) were provided by Lonza (Walkersville, MD) and grown in BEM media supplemented with the BEGM bullet kit (Lonza). NHBE were used for functional studies, and for generation of polarized human airway epithelial cultures (HAE). To generate HAE, NHBE from individual donors were expanded on plastic to generate passage 1 cells, which were subsequently plated (5x104 cells/well) on rat-tail collagen type 1-coated permeable transwell membrane supports (6.5mm; Corning Inc). HAE cultures were grown in B-ALI medium supplemented with inducer (Lonza Inc.) at each media change with provision of an air-liquid interface for approximately 6 weeks to form differentiated, polarized cultures that resemble in vivo pseudostratified mucociliary epithelium.
Primary human normal airway tracheobronchial epithelial cells from de-identified donors (NHBE) (sex: male and female) were provided by Lonza (Walkersville, MD) and grown in BEM media supplemented with the BEGM bullet kit (Lonza). NHBE were used for functional studies, and for generation of polarized human airway epithelial cultures (HAE). To generate HAE, NHBE from individual donors were expanded on plastic to generate passage 1 cells, which were subsequently plated (5x104 cells/well) on rat-tail collagen type 1-coated permeable transwell membrane supports (6.5mm; Corning Inc). HAE cultures were grown in B-ALI medium supplemented with inducer (Lonza Inc.) at each media change with provision of an air-liquid interface for approximately 6 weeks to form differentiated, polarized cultures that resemble in vivo pseudostratified mucociliary epithelium.
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