airway epithelial cell basal media  (ATCC)

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: Clinical Cancer Research

Article Title: Aberrant Activation of Wound-Healing Programs within the Metastatic Niche Facilitates Lung Colonization by Osteosarcoma Cells

doi: 10.1158/1078-0432.CCR-24-0049

Figure Lengend Snippet: Metastasis-associated epithelial cells acquire a chronic wound phenotype. A, Uniform Manifold Approximation and Projection (UMAP) plot of murine alveolar epithelial cells in healthy lung compared with metastasis. The gray circle denotes marked enrichment of DATP in metastasis. B, Dot plot demonstrating expression of key marker genes (columns) in alveolar epithelial cell clusters (row). C, UMAP plot of normal human lung specimens compared with patient lung metastasis samples. The gray circle denotes enrichment of basaloid cells in osteosarcoma metastasis. D, Dot plot demonstrating expression of key marker genes. E, Visium spatial transcriptomic analysis illustrating localization of alveolar epithelial cell subsets enriched in a metastasis-bearing murine lung. F, IHC of Spc (red, AEC2 marker) and Ki67 (green, proliferation marker). L, unaffected lung; T, metastasis. The dash circle denotes lung–metastasis border. Rectangle marks inset shown below. Areas demonstrate examples of Spc + Ki67 + cells. Nuclei are counterstained with DAPI (blue). Scale bar, 50 μm. G, Quantification of AEC2 proliferation. Approximately 100 cells/animal, n = 4. Welch t test: F420 P = 0.0003, K7M2 P ≤ 0.0001, OS-17 P = 0.0004. H, IHC of Krt8 (green, DATP marker). Scale bar, 50 μm. The dash circle denotes lung–metastasis border. Areas demonstrate examples of Krt8 + cells around growing metastatic lesions. Nuclei are counterstained with DAPI (blue). Scale bar, 50 μm. I, Quantification of DATP accumulation (by Krt8 + fluorescence). N = 4 animals, at least 10 regions of interest per animal for metastasis and unaffected lung. Welch t test: P ≤ 0.0001.

Article Snippet: HBEC cells were propagated in human airway epithelial media (ATCC, PCS-300-030) with the Bronchial Epithelial Cell Growth Kit (ATCC, PCS-300-040).

Techniques: Expressing, Marker, Fluorescence

Journal: Clinical Cancer Research

Article Title: Aberrant Activation of Wound-Healing Programs within the Metastatic Niche Facilitates Lung Colonization by Osteosarcoma Cells

doi: 10.1158/1078-0432.CCR-24-0049

Figure Lengend Snippet: Lung epithelial cells induce metastasis-specific osteosarcoma fibronectin matrix formation. A, IHC for fibronectin in several osteosarcoma metastasis models. B, IHC comparing fibronectin staining in tibial tumors versus lung metastasis. C, IHC for fibronectin (green) and Krt8 (red, DATP cells). Dotted box denotes inset, which demonstrates area of metastasis enriched in Krt8 + DATP cells and human (osteosarcoma) fibrillar fibronectin. D, Immunofluorescence of fibronectin matrix in OS-17 cells cultured alone or in co-culture (CoCx) with human bronchial epithelial cells (HBEC; KRT8 = red ). E, Immunofluorescence of fibronectin in OS-17 cells cultured at steady state or stimulated with conditioned HBEC media. Scale bar, 50 μm. Nuclei are counterstained with DAPI (blue).

Article Snippet: HBEC cells were propagated in human airway epithelial media (ATCC, PCS-300-030) with the Bronchial Epithelial Cell Growth Kit (ATCC, PCS-300-040).

Techniques: Staining, Immunofluorescence, Cell Culture, Co-Culture Assay