bronchial epithelial cell growth medium  (PromoCell)

Journal: bioRxiv

Article Title: SenCat: Cataloging human cell senescence through multiomic profiling of multiple senescent primary cell types

doi: 10.64898/2026.02.05.703986

Figure Lengend Snippet: (A) Schematic of the experimental design for induction of systemic senescence with doxorubicin and sample collection across time points. (B) Cluster identities assigned to doxorubicin-treated lung samples profiled by snRNA-seq. (C) UMAP projection of lung samples colored by cluster identity. (D) UMAP projections of senescent cells in lung samples across time points. (E) Bar plot showing the number of senescent cells from lung per cluster and condition. (F,G) GSEA plots of gene set association scores for p53, EMT, NF-κB, Apoptosis, and Hypoxia hallmark pathways (F) and senescence signature lists SenePy, SeneSig, SenMayo, hUSI (G); in fibroblast, epithelial, and endothelial clusters on Day 6. (H) Schematic of the analysis pipeline applied to published aging lung snRNA-seq datasets. (I) UMAP projection of aging lung samples showing senescent cell distribution by age group. (J) Schematic of ligand-receptor inference analysis between senescent fibroblasts and non-senescent epithelial cells. (K) Chord diagram displaying ligand-receptor interactions between senescent fibroblasts (sender cells) and non-senescent epithelial cells (receiver cells) inferred through CellPhoneDB. (L) Dot plot showing expression of the specified ligands across fibroblast clusters at 23 months of age. (M) Schematic and GSEA plot explaining and displaying TGFβ signaling pathway increase during aging in non-senescent epithelial cells at 23 months versus 3 months. See also Figures S11-13 .

Article Snippet: HSAEC lung epithelial cells (ATCC, PCS-301-010) were cultured using an Airway Epithelial Cell Basal Medium plus Bronchial Epithelial Cell Growth Kit (ATCC).

Techniques: Expressing

Journal: Proceedings of the National Academy of Sciences of the United States of America

Article Title: Glycan recognition by collectin-11 drives SARS-CoV-2 infectivity and membrane injury of respiratory epithelial cells

doi: 10.1073/pnas.2521209122

Figure Lengend Snippet: SARS-CoV-2 opsonized with CL-11 drives increased infectivity of respiratory epithelial cells. ( A and B ), Enhancement of BEAS-2B cell infection with SARS-CoV-2 variants pretreated with rCL-11 (0.1, 1.0, and 10.0 µg/mL) or media for 1 h at 37 °C. ( A ) England/02/2020 (MOI 0.005) and ( B ) B.1.617.2 (MOI 0.05). ( C and D ), Enhancement of Calu-3 cell infection with SARS-CoV-2 variants pretreated with rCL-11 (0.1, 1.0, and 10.0 µg/mL) or media for 1 h at 37 °C. ( C ) England/02/2020 (MOI 0.05) and ( D ) B.1.617.2 (MOI 0.05). Virus titers in the supernatant after 24 h determined by plaque assay. Data are pooled from two ( A , C , D ) or three ( B ) independent experiments and represent the mean ± SEM of triplicate or quadruplicate biological samples. Each group compared to 0 µg/mL rCL-11 by one-way ANOVA with Dunnett’s multiple comparison test, * P < 0.05, and *** P < 0.001. ( E and F ), Representative images of crystal violet stained plaque assay plates of BEAS-2B or Calu-3 cells infected with either England/02/2020 ( E ) or B.1.617.2 ( F ) or opsonized with 10 mg/mL rCL-11. ( G ) L-fucose inhibits the enhancement of SARS-CoV-2 (England/02/2020) infectivity driven by CL-11 in BEAS-2B cells. England/02/2020 (MOI 0.05) pretreated with media or with rCL-11 (10.0 µg/mL) in the presence or absence of L-fucose or D-galactose (1- or 10 mM final concentration) and virus titers determined as above. ( H ) Infectivity of SARS-CoV-2 (England/02/2020) opsonized with CL-11 is not inhibited by NHS. Quantification of SARS-CoV-2 England/02/2020 in the supernatant of BEAS-2B cells 24 h after infection with virus (MOI 0.005) pretreated with media or with rCL-11 (10.0 µg/mL) spiked with or without 10% NHS as detailed above. ( G and H ) Data are representative of two independent experiments with the mean ± SEM of five ( G ) and at least six ( H ) biological samples. ** P < 0.01 and *** P < 0.001 determined by one-way ANOVA with Tukey’s multiple comparison test.

Article Snippet: Donor 1: female, 62 y, Caucasian Donor 2: male, 71 y, Caucasian Donor 3: male, 62 y, Caucasian Cryopreserved HBECs from healthy donors obtained from Epithelix and PromoCell were expanded and then seeded (2.12 × 10 5 cells/cm 2 ) onto collagen coated (30 μg/mL) tranwells and cultured submerged in Bronchial Epithelial Cell Growth Medium (Promocell) until confluent.

Techniques: Infection, Virus, Plaque Assay, Comparison, Staining, Concentration Assay

Journal: Molecular Therapy. Nucleic Acids

Article Title: ACE-2 blockade and TMPRSS2 inhibition mitigate SARS-CoV-2 severity following cigarette smoke exposure in airway epithelial cells in vitro

doi: 10.1016/j.omtn.2025.102580

Figure Lengend Snippet: ACE-2 and TMPRSS2 expression are elevated in cigarette-smoke-exposed ferret lung (A) Dot plot shown from RNA-seq analysis of SARS-CoV-2 receptor and associated gene expression in lung tissue of ferrets exposed to cigarette smoke for 6 months. (B) ACE-2 and (C) TMPRSS2 mRNA expression in lung tissue from smoke-exposed vs. air control ferrets as assessed by real-time qPCR. (D) Representative western blot and (E) quantification of ACE-2 expression in smoke-exposed ferret lung. (F) Quantification of TMPRSS2 expression in smoke-exposed ferret lung. (G) Representative immunofluorescence images of smoke-exposed and air-control ferret lung sections stained with ACE-2 antibody (red) and DAPI (blue). Scale bars, 50 μM. Areas of magnification (magnification 40X) are outlined by the red dashed line. (H) Representative immunofluorescence images of smoke-exposed and air-control ferret lung sections stained with TMPRSS-2 antibody (Green), Muc5AC (red), and DAPI (blue). Scale bars, 50 μM. Areas of magnification (magnification 40X) are outlined by the red dashed line. Real-time qPCR analysis of (I) ACE2 and (J) TMPRSS2 mRNA expression in terminally differentiated ferret tracheal epithelial cells (FTECs) exposed to cigarette smoke extract (CSE) or vehicle control. ∗ p < 0.05, ∗∗ p < 0.005,∗∗∗ p < 0.0005.

Article Snippet: Primary HBE cells were isolated from lung explants at the time of organ transplantation from COPD patients, healthy donors without lung disease, and healthy donors, expanded in submerged culture for one or two passages in bronchial epithelial growth medium (BEGM, Lonza, Walksville, MD), and then seeded on Transwell membranes (Corning, New York, NY) as described previously., , Cells were grown at ALI in PneumaCult-ALI media (Stem Cell Technology) for 3–4 weeks until terminally differentiated.

Techniques: Expressing, RNA Sequencing, Gene Expression, Control, Western Blot, Immunofluorescence, Staining

Journal: Molecular Therapy. Nucleic Acids

Article Title: ACE-2 blockade and TMPRSS2 inhibition mitigate SARS-CoV-2 severity following cigarette smoke exposure in airway epithelial cells in vitro

doi: 10.1016/j.omtn.2025.102580

Figure Lengend Snippet: ACE-2 and TMPRSS2 expression are elevated in CSE-exposed human airway cells Real-time qPCR measurement of (A) ACE-2 and (B) TMPRSS2 mRNA expression in Calu-3 cells exposed to cigarette smoke extract (CSE) or vehicle control for 48 h Real-time qPCR measurement of (C) ACE-2 and (D) TMPRSS2 mRNA expression in primary human bronchial epithelial (HBE) cells derived from healthy control donors and treated with CSE or vehicle control for 48 h. N = 3 monolayers/condition across 3–4 different donors. Western blot image HBE cells treated with cigarette smoke extract for 24 h (E) and quantitation of western blot for ACE2 (F) and TMPRSS2 (G). Real-time qPCR measurement of (H) ACE-2 and (I) TMPRSS2 mRNA expression in bronchial epithelial cells derived from patients with COPD (COPD HBE) or healthy non-smoker donors. N = 3 monolayers/condition across 3–4 different donors. Western blot analysis of lung tissue from COPD and healthy non-smoker donors (J), with corresponding quantification of TMPRSS2 protein expression (K). (G) Representative western blot for ACE2 (L), with quantification of ACE2 protein expression comparing non-smokers vs. COPD (M) and non-smokers vs. smokers (N). N = 3–4 monolayers/conditions derived from 3 to 4 different donors. ∗ p < 0.05, ∗∗ p < 0.005.

Article Snippet: Primary HBE cells were isolated from lung explants at the time of organ transplantation from COPD patients, healthy donors without lung disease, and healthy donors, expanded in submerged culture for one or two passages in bronchial epithelial growth medium (BEGM, Lonza, Walksville, MD), and then seeded on Transwell membranes (Corning, New York, NY) as described previously., , Cells were grown at ALI in PneumaCult-ALI media (Stem Cell Technology) for 3–4 weeks until terminally differentiated.

Techniques: Expressing, Control, Derivative Assay, Western Blot, Quantitation Assay

Journal: Molecular Therapy. Nucleic Acids

Article Title: ACE-2 blockade and TMPRSS2 inhibition mitigate SARS-CoV-2 severity following cigarette smoke exposure in airway epithelial cells in vitro

doi: 10.1016/j.omtn.2025.102580

Figure Lengend Snippet: SARS-CoV-2 infection is increased in CSE-exposed FTECs and Calu-3 cells (A) Schematic outline for experiments evaluating the relationship between cigarette smoking and SARS-CoV-2 infection in ferret tracheal epithelial cells (FTECs) and Calu-3 cells. Cells were treated with cigarette smoke extract (CSE) or vehicle control for 48 h prior to inoculation with SARS-CoV-2 (MOI-3). After 72 h of SARS-CoV-2 infection with concomitant CSE exposure, cells were harvested for analysis. (B) Quantification of viral load by real-time qPCR of SARS-CoV-2 mRNA at 3 days post-infection in FTECs exposed to CSE or vehicle control, with (C) visualization and (D) quantification of SARS-CoV-2-infected cells by foci forming assay in Vero-E6 cells. N = 3–5 well per conditions. (E) Quantification of viral load by real-time qPCR at 3 days after SARS-CoV-2 infection in Calu-3 cells exposed to CSE or vehicle control, with (F) visualization and (G) quantification of SARS-CoV-2-infected cells by foci forming assay in Vero-E6 cells. N = 3–5 well per conditions. ∗ p < 0.05, ∗∗ p < 0.01.

Article Snippet: Primary HBE cells were isolated from lung explants at the time of organ transplantation from COPD patients, healthy donors without lung disease, and healthy donors, expanded in submerged culture for one or two passages in bronchial epithelial growth medium (BEGM, Lonza, Walksville, MD), and then seeded on Transwell membranes (Corning, New York, NY) as described previously., , Cells were grown at ALI in PneumaCult-ALI media (Stem Cell Technology) for 3–4 weeks until terminally differentiated.

Techniques: Infection, Control

Journal: Molecular Therapy. Nucleic Acids

Article Title: ACE-2 blockade and TMPRSS2 inhibition mitigate SARS-CoV-2 severity following cigarette smoke exposure in airway epithelial cells in vitro

doi: 10.1016/j.omtn.2025.102580

Figure Lengend Snippet: SARS-CoV-2 genomic replication is increased in CSE-exposed normal HBE cells and HBE cells derived from COPD patients (A) Transepithelial electrical resistance (TEER) measurements in vehicle-treated, cigarette smoke extract (CSE)-treated, and COPD human bronchial epithelial (HBE) monolayer cultures infected with SARS-CoV-2 or mock infection. The data were obtained by averaging three independent Transwell reads, each of which represented the mean of three separate readings. ( N = 3–5) HBE. Each line corresponds to a distinct donor (3–5 donors in total), and each donor is represented by 3–5 replicates. (B) Quantification of viral load by real-time qPCR of SARS-CoV-2 mRNA at 72 h post-infection with SARS-CoV-2 in CSE- or vehicle-exposed HBE cells. N = 3 different donors, with each line representing a separate donor. (C) Quantification of viral load by real-time qPCR of SARS-CoV-2 mRNA at 72 h post-infection with SARS-CoV-2 in healthy non-smoker or COPD HBE cells. N = monolayers/condition derived from 3 to 5 different donors. (D and E) Representative images using RNAscope in situ hybridization for comparable detection of genomic RNA with only the SARS-CoV-2-specific S probe. Images show CSE- or vehicle-exposed HBE cells, and healthy non-smoker or COPD HBE cells, at 72 h post-inoculation with (D) mock infection or (E) SARS-CoV-2. SARS-CoV-2 (red), TMPRSS2 (green), ACE-2 (white), nuclei (blue). ∗∗ p < 0.01.

Article Snippet: Primary HBE cells were isolated from lung explants at the time of organ transplantation from COPD patients, healthy donors without lung disease, and healthy donors, expanded in submerged culture for one or two passages in bronchial epithelial growth medium (BEGM, Lonza, Walksville, MD), and then seeded on Transwell membranes (Corning, New York, NY) as described previously., , Cells were grown at ALI in PneumaCult-ALI media (Stem Cell Technology) for 3–4 weeks until terminally differentiated.

Techniques: Derivative Assay, Infection, RNAscope, In Situ Hybridization

Journal: Molecular Therapy. Nucleic Acids

Article Title: ACE-2 blockade and TMPRSS2 inhibition mitigate SARS-CoV-2 severity following cigarette smoke exposure in airway epithelial cells in vitro

doi: 10.1016/j.omtn.2025.102580

Figure Lengend Snippet: Simultaneous ACE2 blockade and TMPRSS2 inhibition reduces SARS-CoV-2 infection after CSE exposure in FTECs (A) Scheme depicting the approach to ACE2 antisense oligonucleotide (ASO) treatment, cigarette smoke extract (CSE) exposure, and SARS-CoV-2 infection in ferret tracheal epithelial cells (FTECs). Seven days of treatment with ACE2 ASO (20 μM) or control ASO was followed by a 48-h incubation period in CSE or vehicle control prior to inoculation with SARS-CoV-2 or no virus for 72 h of infection. Assessment of mRNA expression of (B) ACE-2 and (C) TMPRSS2 following the scheme depicted in (A). (D) Scheme depicting the approach to ACE2 ASO and camostat mesylate treatment, CSE exposure, and SARS-CoV-2 infection in FTECs. Camostat mesylate (100 mM) was added for 2 h on day 9, prior to inoculation with SARS-CoV-2. Assessment of (E) viral load and mRNA expression of (F) ACE-2 and (G) SARS-Cov2 infection for a shorter duration following the scheme depicted in (D). Viral load following shorter duration of infection (H) and (I). Real-time qPCR was used for mRNA quantification in all studies. ∗ p < 0.05, ∗∗ p < 0.01, ∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001.

Article Snippet: Primary HBE cells were isolated from lung explants at the time of organ transplantation from COPD patients, healthy donors without lung disease, and healthy donors, expanded in submerged culture for one or two passages in bronchial epithelial growth medium (BEGM, Lonza, Walksville, MD), and then seeded on Transwell membranes (Corning, New York, NY) as described previously., , Cells were grown at ALI in PneumaCult-ALI media (Stem Cell Technology) for 3–4 weeks until terminally differentiated.

Techniques: Inhibition, Infection, Control, Incubation, Virus, Expressing

Journal: International Journal of Molecular Medicine

Article Title: MEIS1-regulated miR-488-3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C

doi: 10.3892/ijmm.2025.5583

Figure Lengend Snippet: miR-488-3p overexpression inhibits LSCC cell proliferation, migration, invasion and epithelial-mesenchymal transition. (A) Quantitative PCR analysis of miR-488-3p expression in LSCC cells transfected with miR-488-3p or NC mimics. (B) Cell Counting Kit-8 and (C) colony formation assays detected the proliferative ability of LSCC cells transfected with miR-488-3p or NC mimics. Transwell assays determined the (D) migration and (E) invasion of LSCC cells transfected with miR-488-3p or NC mimics. (F) Western blotting of the protein levels of E-cadherin, N-cadherin and vimentin. Cropped images represent different blots, but samples are from the same experiment and the blots were processed in parallel. Scale bars, 100 µ m. * P<0.05, ** P<0.01 and *** P<0.001. miR, microRNA; LSCC, laryngeal squamous cell carcinoma; NC, negative control.

Article Snippet: FD-LSC-1 and 2BS cells were cultured in Bronchial Epithelial Cell Growth Medium (Lonza Group, Ltd.) and MEM (Procell Life Science &Technology Co., Ltd.) with 10% FBS, respectively.

Techniques: Over Expression, Migration, Real-time Polymerase Chain Reaction, Expressing, Transfection, Cell Counting, Western Blot, Negative Control

Journal: International Journal of Molecular Medicine

Article Title: MEIS1-regulated miR-488-3p suppresses the malignant progression of laryngeal squamous cell carcinoma by targeting ACVR1C

doi: 10.3892/ijmm.2025.5583

Figure Lengend Snippet: ACVR1C knockdown inhibits LSCC cell proliferation, migration, invasion and epithelial-mesenchymal transition. (A) Analysis of ACVR1C expression in RNA-seq data ( GSE127165 ). (B) Quantitative PCR and (C) western blot analysis of the mRNA and protein expression levels of ACVR1C in AMC-HN-8 and FD-LSC-1 cells transfected with si-ACVR1C-1 and 2 or si-NC. Cropped images represent different blots, but samples are from the same experiment and the blots were processed in parallel. (D) Cell Counting Kit-8 and (E) colony formation assays investigated the proliferative ability of LSCC cells transfected with si-ACVR1C or si-NC. In the Transwell assay, ACVR1C knockdown significantly suppressed the (F) migration and (G) invasion of LSCC cells. (H) Western blot analysis of the protein levels of E-cadherin, N-cadherin and vimentin in LSCC cells transfected with si-ACVR1C or si-NC. Cropped images represent different blots, but samples are from the same experiment and the blots were processed in parallel. Scale bar, 100 µ m. * P<0.05, ** P<0.01 and *** P<0.001. ACVR1C, activin A receptor type 1C; LSCC, laryngeal squamous cell carcinoma; NC, negative control; si, small interfering RNA.

Article Snippet: FD-LSC-1 and 2BS cells were cultured in Bronchial Epithelial Cell Growth Medium (Lonza Group, Ltd.) and MEM (Procell Life Science &Technology Co., Ltd.) with 10% FBS, respectively.

Techniques: Knockdown, Migration, Expressing, RNA Sequencing, Real-time Polymerase Chain Reaction, Western Blot, Transfection, Cell Counting, Transwell Assay, Negative Control, Small Interfering RNA

Journal: bioRxiv

Article Title: Studying a human genetic model of lung squamous cell carcinoma with organotypic cultures and xenografts uncovers distinct advantages of each system and implicates NOTCH1 loss in tumour development

doi: 10.1101/2025.08.28.672595

Figure Lengend Snippet: Comparative histological analysis of xenografts and ALI cultures generated with mutant hBECs from the same donor. A. Representative H&E images of the epithelial structure of xenografts. Images were taken from areas that comprise surrounding mouse stroma, epithelial layer and lumen for xenografts with cystic structure. For solid xenografts, images were taken from areas that comprise surrounding mouse stroma and tumour nests. For TC+P and TC+PS mutants, representative images of areas with squamous and mucociliary morphologies are shown separately. L=Lumen; d= dyskeratosis. B. Representative H&E images of the epithelial structure of ALI cultures. C. Representative images of p63 immunohistochemical staining of xenografts. L=Lumen. D. Images depicting the presence of intercellular bridges and keratin pearls in TC+PKS mutants, two features of well-differentiated LUSC. Arrows mark the presence of intercellular bridges. kp= keratin pearl. E. TTF-1 immunohistochemical staining of a TC+PKS xenograft showing the absence of expression of this lung adenocarcinoma marker. F . Quantification of total invading single cells into the mouse stroma and an example image of an invading single cell stained for human mitochondria. Data shown as mean±SEM (n=6 xenografts). Adjusted p-values were calculated using one-way ANOVA followed by Tukey’s multiple comparisons test (only significant comparisons are shown) G. Images depicting a xenograft area with adjacent mucociliary and squamous morphology in a TC+PS mutant. Images show H&E staining, and immunohistochemical staining for mCherry, acetylated-tubulin and MUC5AC. Areas with mucociliary differentiation show expression of acetylated-tubulin (cilia) and MUC5AC (goblet cells). Mc=mucociliary; sq=squamous. Statistical significance shown as: ∗p < 0.05, ∗∗p < 0.01

Article Snippet: A total of 30,000 bronchial epithelial cells were pelleted and resuspended in 100 μl of Airway Epithelial Cell Growth Medium (PromoCell, C-21160), then seeded directly onto the collagen-coated membranes, which were placed in individual wells of a 24-well plate.

Techniques: Generated, Mutagenesis, Immunohistochemical staining, Staining, Expressing, Marker