small airway epithelial cell media  (PromoCell)

Journal: bioRxiv

Article Title: Dynamic Culture Improves the Predictive Power of Bronchial and Alveolar Airway Models of SARS-CoV-2 Infection

doi: 10.1101/2025.07.21.665885

Figure Lengend Snippet: (A) Schematic of the PhysioMimix® bronchial MPS, showing human primary bronchial epithelial cells cultured at air–liquid interface (ALI) under static and dynamic flow conditions. (B) Representative H&E-stained histological sections of bronchial tissues after 14 days of differentiation under static or MPS conditions. Scale bar, 50 µm. (C) Trans-epithelial electrical resistance (TEER) measurements over the 14-day ALI differentiation period. (D) Gene expression of Club cell (SCGB1A1) and Goblet cell (MUC5AC) markers in static versus MPS cultures after 14 days. GP = 0.0332 (*), 0.0021 (**). (E) Immunofluorescence staining of MPS bronchial tissue for acetylated α- tubulin (yellow), mucus (MUC5AC, green), actin (phalloidin, magenta), and nuclei (Hoechst 33342, blue). Scale bar, 100 µm.

Article Snippet: HPMECs, SAECs and NHBEs were maintained at 37°C, 5% CO 2 and 95% humidity in cell-type specific media (EGM-2 basal medium bullet kit [Lonza, CC-3162]; small airway epithelial cell media [PromoCell, C-211170]; PneumaCult-Ex Plus medium [STEMCELL Technologies, 05040], respectively) in T75 flasks until they reached 70% confluency.

Techniques: Cell Culture, Staining, Gene Expression, Immunofluorescence

Journal: bioRxiv

Article Title: Dynamic Culture Improves the Predictive Power of Bronchial and Alveolar Airway Models of SARS-CoV-2 Infection

doi: 10.1101/2025.07.21.665885

Figure Lengend Snippet: (A) Schematic of the PhysioMimix® alveolar MPS, showing human primary small airway epithelial cells cultured at air– liquid interface (ALI) under static and dynamic flow conditions. (B) Representative H&E-stained histological sections of alveolar tissues after 14 days of differentiation. Scale bar, 50 µm. (C) Trans-epithelial electrical resistance (TEER) measurements over the 14-day ALI differentiation period. (D) Gene expression of alveolar type I (AT1; AQP5) and alveolar type II (AT2; SFTPB) cell markers in static versus MPS cultures. GP = 0.0021 (**). (E) Immunofluorescence staining of alveolar MPS tissue for AT1 cells (RAGE, green), AT2 cells (SFTPB, yellow), actin (phalloidin, magenta), and nuclei (Hoechst 33342, blue). Scale bar, 100 µm.

Article Snippet: HPMECs, SAECs and NHBEs were maintained at 37°C, 5% CO 2 and 95% humidity in cell-type specific media (EGM-2 basal medium bullet kit [Lonza, CC-3162]; small airway epithelial cell media [PromoCell, C-211170]; PneumaCult-Ex Plus medium [STEMCELL Technologies, 05040], respectively) in T75 flasks until they reached 70% confluency.

Techniques: Cell Culture, Staining, Gene Expression, Immunofluorescence

Journal: bioRxiv

Article Title: Dynamic Culture Improves the Predictive Power of Bronchial and Alveolar Airway Models of SARS-CoV-2 Infection

doi: 10.1101/2025.07.21.665885

Figure Lengend Snippet: (A) Schematic of the PhysioMimix® lung MPS co-culture system. Epithelial cells were seeded on the apical side of the Transwell® insert and human pulmonary microvascular endothelial cells (HPMVECs) on the basolateral side, under air–liquid interface (ALI) and dynamic flow conditions. (B) Representative H&E and Alcian blue-stained histological section of bronchial MPS co-culture after 14 days. Mucus is stained blue; endothelial cells are indicated by white arrows. Scale bar, 50 µm. (C) TEER measurements comparing epithelial monoculture, endothelial monoculture, and epithelial–endothelial co-culture over 14 days under ALI conditions. (D) TEER comparison of bronchial and alveolar co-cultures grown under static or dynamic flow MPS conditions over the 14-day ALI differentiation period. (E) Gene expression of Club cell (SCGB1A1) and Goblet cell (MUC5AC) markers in NHBE cells before culture (NHBE pellet) and after 14 days of differentiation in MPS co- culture. (F) Gene expression of alveolar markers - AT1 (AQP5) and AT2 (SFTPB) - in SAEC cells before culture (SAEC pellet) and after 14 days of MPS co-culture. (G) Immunofluorescence staining of bronchial MPS co-culture tissue for acetylated α-tubulin (yellow), mucus (MUC5AC, green), actin (phalloidin, red), and nuclei (Hoechst 33342, blue). Top row shows epithelial layer; bottom row shows endothelial layer. Scale bar, 100 µm. (H) Immunofluorescence staining of alveolar MPS co-culture tissue after 14 days of differentiation, showing surfactant (SFTPB, green), actin (phalloidin, magenta), and nuclei (Hoechst 33342, blue). Endothelial cells are marked with white arrows. Scale bar, 20 µm.

Article Snippet: HPMECs, SAECs and NHBEs were maintained at 37°C, 5% CO 2 and 95% humidity in cell-type specific media (EGM-2 basal medium bullet kit [Lonza, CC-3162]; small airway epithelial cell media [PromoCell, C-211170]; PneumaCult-Ex Plus medium [STEMCELL Technologies, 05040], respectively) in T75 flasks until they reached 70% confluency.

Techniques: Co-Culture Assay, Staining, Comparison, Gene Expression, Immunofluorescence

Journal: bioRxiv

Article Title: Aberrant Transitional Alveolar Epithelial Cells Promote Pathogenic Activation of Lung Fibroblasts in Preclinical Fibrosis Modeling

doi: 10.1101/2024.06.17.599351

Figure Lengend Snippet: (A) Schematic of chronic tamoxifen-induced Sftpc C121G mutation model and subsequent cell sorting for single-cell RNA sequencing (scRNA-seq). (B) Representative H&E staining with fibrosis severity scoring (normal = blue, moderate = green, and severe = red) and bar graph showing relative amounts of injury per lung lobe. (C) scRNA-seq UMAP representation of mesenchymal, proximal (airway), and distal (alveolar) epithelial subsets in both Sftpc WT and Sftpc C121G mice. Inserts depict annotated subset analysis of distal epithelial and mesenchymal populations. UPR, unfolded protein response. (D) UMAP representation of gene marker scoring of distal epithelial populations. (E) Dot plot of individual gene expression in distal epithelial cell populations. (F) UMAP representation of defined profibrotic genes enriched within the aberrant transitional cell population. (G) Gene score representation by UMAP and dot plots of signature gene markers characterized in murine basaloid-like cells . (H) Gene score dot plots from human aberrant basaloid populations ( , ). (I) Trajectory analysis suggesting aberrant transitional cells are a terminal cell fate in the Sftpc C121G alveolar epithelium.

Article Snippet: Cell suspensions were resuspended in a mix of 50% GFR-Matrigel (Corning) and 50% Small Airway Epithelial Cell Growth Media (SAGM; Lonza) and plated in a 24-well Falcon Cell Culture Insert (Falcon).

Techniques: Mutagenesis, FACS, RNA Sequencing, Staining, Marker, Gene Expression

Journal: bioRxiv

Article Title: Aberrant Transitional Alveolar Epithelial Cells Promote Pathogenic Activation of Lung Fibroblasts in Preclinical Fibrosis Modeling

doi: 10.1101/2024.06.17.599351

Figure Lengend Snippet: (A) Schematic demonstrating single-cell ligand-receptor mapping using NICHES. (B) UMAP representation of unbiased epithelial-to-mesenchymal ligand-receptor mapping clustered dependent on distinct signaling archetypes. (C) UMAPs labeled by epithelial sending cell type and mesenchymal receiving cell type populations. Ratios of each cell type within a given cluster is provided. (D) Heatmap of individual ligand-receptor pairs that define each cluster archetype. (E) Representative UMAPs of known homeostatic and profibrotic ligand-receptor pairs that are altered in fibrosis.

Article Snippet: Cell suspensions were resuspended in a mix of 50% GFR-Matrigel (Corning) and 50% Small Airway Epithelial Cell Growth Media (SAGM; Lonza) and plated in a 24-well Falcon Cell Culture Insert (Falcon).

Techniques: Labeling

Journal: bioRxiv

Article Title: Aberrant Transitional Alveolar Epithelial Cells Promote Pathogenic Activation of Lung Fibroblasts in Preclinical Fibrosis Modeling

doi: 10.1101/2024.06.17.599351

Figure Lengend Snippet: (A) Schematic demonstrating single-cell ligand-receptor mapping using NICHES. (B) UMAP representation of unbiased mesenchymal-to-epithelial ligand-receptor mapping clustered dependent on distinct signaling archetypes. (C) UMAPs labeled by mesenchymal sending cell type and epithelial receiving cell type populations. Ratios of each cell type within a given cluster is provided. (D) Heatmap of individual ligand-receptor pairs that define each cluster archetype. (E) Representative UMAPs of known homeostatic and profibrotic ligand-receptor pairs that are altered in fibrosis.

Article Snippet: Cell suspensions were resuspended in a mix of 50% GFR-Matrigel (Corning) and 50% Small Airway Epithelial Cell Growth Media (SAGM; Lonza) and plated in a 24-well Falcon Cell Culture Insert (Falcon).

Techniques: Labeling

Journal: bioRxiv

Article Title: Aberrant Transitional Alveolar Epithelial Cells Promote Pathogenic Activation of Lung Fibroblasts in Preclinical Fibrosis Modeling

doi: 10.1101/2024.06.17.599351

Figure Lengend Snippet: (A) Previously published UMAP representation of epithelial cell types captured by scRNAseq at 18 time points (Strunz et al., 2020), and re-clustering of the alveolar epithelium. (B) UMAP representation and (C) Dot plot gene scores according to alveolar epithelium cell clusters. (D) Schematic tamoxifen (TMX) dosing and bleomycin administration in Sftpc lineage traced mice prior to Day 14 analysis. (E) IFC demonstrating tdT + CD44+ elongated cells at Day 14 in bleomycin model. Scale bar = 20 µm (F) Flow cytometry analysis of CD44 upregulation in mice that received bleomycin at Day 14. Student’s t-test, **** P<0.0001. (G) Gene expression analysis of total healthy tdT + AT2s compared to CD44 - and CD44 + tdT + cells at Day 14 in mice that received bleomycin. (H) Schematic of healthy alveolar fibroblast exposure to conditioned supernatant from AT2s 2D cultured for five days. (I) Gene expression of alveolar fibroblasts after two days of supernatant exposure. Statistical analysis by one-way ANOVA (*p < 0.05).

Article Snippet: Cell suspensions were resuspended in a mix of 50% GFR-Matrigel (Corning) and 50% Small Airway Epithelial Cell Growth Media (SAGM; Lonza) and plated in a 24-well Falcon Cell Culture Insert (Falcon).

Techniques: Flow Cytometry, Gene Expression, Cell Culture

Journal: bioRxiv

Article Title: Aberrant Transitional Alveolar Epithelial Cells Promote Pathogenic Activation of Lung Fibroblasts in Preclinical Fibrosis Modeling

doi: 10.1101/2024.06.17.599351

Figure Lengend Snippet: (A) Schematic depicting mixed organoid cultures derived from various combinations of AT2/transitional cells and bulk Pdgfra + mesenchymal cells from Sftpc WT and Sftpc C121G mice. (B) Representative imaging of organoid cultures. Scale bar = 1mm. (C) Colony forming efficiency (CFE; %) and (D) organoid diameter (µm) and of organoids at Day 14 in culture. (E) Schematic depicting EpCAM+ magnetic bead sorting from Day 14 organoids prior to gene expression analysis. (F) Gene expression analysis from enriched epithelial or fibroblast populations derived from Day 14 organoids. Ordinary one-way ANOVA with uncorrected Fisher’s LSD multiple comparisons were performed. (*) were used when comparing samples cultured within the same fibroblast condition. (#) were used when comparing samples within the same epithelial condition. */# P<0.5, **/## P<0.01, ***/### P<0.001, ****/#### P<0.0001.

Article Snippet: Cell suspensions were resuspended in a mix of 50% GFR-Matrigel (Corning) and 50% Small Airway Epithelial Cell Growth Media (SAGM; Lonza) and plated in a 24-well Falcon Cell Culture Insert (Falcon).

Techniques: Derivative Assay, Imaging, Gene Expression, Cell Culture