bebm bronchial epithelial cell growth basal medium  (Lonza)

Journal: Bioengineering

Article Title: A 3D Epithelial–Mesenchymal Co-Culture Model of the Airway Wall Using Native Lung Extracellular Matrix

doi: 10.3390/bioengineering11090946

Figure Lengend Snippet: Schematic representation of the model assembly steps. Workflow for the preparation and analysis of the 3D co-culture of primary human fibroblasts and epithelial cells with lung ECM hydrogels. Created with biorender.com.

Article Snippet: Primary human lung epithelial cells ( n = 5) were cultured in Bronchial Epithelial Cell Growth Basal Medium (BEBM, Lonza) supplemented with the Bronchial Epithelial SingleQuotsTM Kit (Lonza) (BEGM).

Techniques: Co-Culture Assay

Journal: Bioengineering

Article Title: A 3D Epithelial–Mesenchymal Co-Culture Model of the Airway Wall Using Native Lung Extracellular Matrix

doi: 10.3390/bioengineering11090946

Figure Lengend Snippet: Model assembly with porcine lung ECM hydrogel and its contraction over time. ( A ) Macroscopic view of the 10 and 20 mg/mL porcine lung ECM hydrogel 3D co-culture model: side view directly after full assembly; top view after 4 days submerged culture; 7-day ALI culture; and 14-day ALI culture. ( B ) Microscopic (20×) images after initial seeding of epithelial cells on and fibroblast seeding in ECM hydrogel showing reduced visibility after hydrogel contraction. ( C ) Brightfield and fluorescent microscopic images (10×): images 1 and 5 days after epithelial cell seeding on top of fibroblast-containing or empty 10 mg/mL porcine lung ECM hydrogels. Cells are stained with DAPI to visualize nuclei. ECM: extracellular matrix, ALI: air–liquid interface, FN: fibronectin, BSA: bovine serum albumin, DAPI: 4′,6-diamidino-2-phenylindole.

Article Snippet: Primary human lung epithelial cells ( n = 5) were cultured in Bronchial Epithelial Cell Growth Basal Medium (BEBM, Lonza) supplemented with the Bronchial Epithelial SingleQuotsTM Kit (Lonza) (BEGM).

Techniques: Co-Culture Assay, Staining

Journal: Bioengineering

Article Title: A 3D Epithelial–Mesenchymal Co-Culture Model of the Airway Wall Using Native Lung Extracellular Matrix

doi: 10.3390/bioengineering11090946

Figure Lengend Snippet: Different porcine lung ECM hydrogel contraction outcomes. Haematoxylin and eosin-stained porcine lung ECM hydrogel co-culture model after 14 days of air–liquid interface culture. ( A ) Epithelial confluency spanning from the hydrogel across the membrane. ( B ) Epithelial confluence covering the entire contracted ECM and small enveloped accumulated cell pockets (arrow). ( C ) Folding of the entire hydrogel during contraction. Scale bars are 500 µm. ECM: extracellular matrix.

Article Snippet: Primary human lung epithelial cells ( n = 5) were cultured in Bronchial Epithelial Cell Growth Basal Medium (BEBM, Lonza) supplemented with the Bronchial Epithelial SingleQuotsTM Kit (Lonza) (BEGM).

Techniques: Staining, Co-Culture Assay, Membrane

Journal: Bioengineering

Article Title: A 3D Epithelial–Mesenchymal Co-Culture Model of the Airway Wall Using Native Lung Extracellular Matrix

doi: 10.3390/bioengineering11090946

Figure Lengend Snippet: Histological analysis of primary airway epithelial cell differentiation prior to and after air–liquid interface (ALI) culture. Tubulin immunostaining (ciliated cells), Alcian blue staining (goblet cells), MUC5AC immunostaining (club cells), and KRT5 immunostaining (basal cells). Submerged culture was fixed after epithelial cells reached confluence, whereas ALI culture exposed the submerged cultured cells to air for 14 days before fixation. ( A ) Intact control human lung. ( B ) Representative images of sections (40× magnification) of submerged- and ALI-cultured airway epithelial cells on top of a collagen I–fibronectin–bovine serum albumin-coated membrane. ( C ). Quantified differentiated epithelial cells on membranes coated with collagen I-FN-BSA. ( D ). Representative images of sections (40× magnification) of submerged- and ALI-cultured airway epithelial cells on top of a collagen IV-coated membrane. ( E ) Quantified differentiated epithelial cells on membranes coated with collagen IV. Scale bars are 50 µm. All data are shown as means with all available data plotted individually ( n = 5). FN: fibronectin, BSA: bovine serum albumin, ALI: air–liquid interface, MUC5AC: mucin-5AC, KRT5: keratin 5. Made with Biorender.com.

Article Snippet: Primary human lung epithelial cells ( n = 5) were cultured in Bronchial Epithelial Cell Growth Basal Medium (BEBM, Lonza) supplemented with the Bronchial Epithelial SingleQuotsTM Kit (Lonza) (BEGM).

Techniques: Cell Differentiation, Immunostaining, Staining, Cell Culture, Control, Membrane

Journal: Bioengineering

Article Title: A 3D Epithelial–Mesenchymal Co-Culture Model of the Airway Wall Using Native Lung Extracellular Matrix

doi: 10.3390/bioengineering11090946

Figure Lengend Snippet: Histological analysis of primary airway epithelial, fibroblast, and porcine lung ECM co-culture model of cell differentiation prior to and after air–liquid interface (ALI) culture. Tubulin immunostaining (ciliated cells), Alcian blue staining (goblet cells), MUC5AC immunostaining (club cells), and KRT5 immunostaining (basal cells). Submerged culture was fixed after epithelial cells reached confluence, whereas ALI culture exposed the submerged cultured cells to air for 14 days before fixation. ( A ) Representative images of sections (40× magnification) of submerged- and ALI-cultured porcine ECM (conc. 10 mg/mL) epithelial cell fibroblast co-culture model. ( B ). Quantified differentiated epithelial cells on membranes coated with collagen I-FN-BSA. ( C ) Representative images of sections (40× magnification) of porcine ECM (conc. 20 mg/mL) epithelial cell fibroblast co-culture model. ( D ) Quantified differentiated epithelial cells on membranes coated with collagen IV. Scale bars are 50 µm. All data are shown as means with all available data plotted individually ( n = 1–5). ECM: extracellular matrix, ALI: air–liquid interface, MUC5AC: mucin-5AC, KRT5: keratin 5. Made with Biorender.com.

Article Snippet: Primary human lung epithelial cells ( n = 5) were cultured in Bronchial Epithelial Cell Growth Basal Medium (BEBM, Lonza) supplemented with the Bronchial Epithelial SingleQuotsTM Kit (Lonza) (BEGM).

Techniques: Co-Culture Assay, Cell Differentiation, Immunostaining, Staining, Cell Culture

Journal: Bioengineering

Article Title: A 3D Epithelial–Mesenchymal Co-Culture Model of the Airway Wall Using Native Lung Extracellular Matrix

doi: 10.3390/bioengineering11090946

Figure Lengend Snippet: Histological analysis of primary airway epithelial, fibroblast, and human lung ECM (HECM) co-culture model of cell differentiation prior to and after air–liquid interface (ALI) culture. Tubulin immunostaining (ciliated cells), Alcian blue staining (goblet cells), MUC5AC immunostaining (club cells), and KRT5 immunostaining (basal cells). Submerged culture was fixed after epithelial cells reached confluence, whereas ALI culture exposed the submerged-cultured cells to air for 14 days before fixation. ( A ) Representative images of sections (40× magnification) of submerged- and ALI-cultured control human lung ECM hydrogel epithelial cell fibroblast co-culture model. ( B ). Quantified differentiated epithelial cells on membranes coated with collagen I-FN-BSA. ( C ). Representative images of sections (40× magnification) of submerged- and ALI-cultured COPD human lung ECM hydrogel epithelial cell fibroblast co-culture model. ( D ) Quantified differentiated epithelial cells on membranes coated with collagen IV. Scale bars are 50 µm. All data are shown as means with all available data plotted individually ( n = 5). ALI: air–liquid interface, MUC5AC: mucin-5AC, KRT5: keratin 5. Made with Biorender.com.

Article Snippet: Primary human lung epithelial cells ( n = 5) were cultured in Bronchial Epithelial Cell Growth Basal Medium (BEBM, Lonza) supplemented with the Bronchial Epithelial SingleQuotsTM Kit (Lonza) (BEGM).

Techniques: Co-Culture Assay, Cell Differentiation, Immunostaining, Staining, Cell Culture, Control