bronchial epithelial cell growth medium  (PromoCell)

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: 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

Journal: bioRxiv

Article Title: The RNA binding protein Insulin Growth factor 2 binding Protein 3 (IGF2BP3) modulates IL-13/IL-4 signalling in human bronchial epithelial cells and is dysregulated in type 2 disease

doi: 10.1101/2025.07.19.665669

Figure Lengend Snippet: IGF2BP3 is increased in type 2 disease. A. Left: normalized expression levels of IGF2BP3 mRNA in severe asthma (SA) vs healthy control (HC) primary BECs at total and polyribosome-bound levels; centre: IGF2BP3 normalized microarray expression data comparing airway epithelial cells from healthy non-atopic (HNA) and atopic asthma (AA) children; right IGF2BP3 normalized microarray expression data comparing nasal lavage samples from children during Picornavirus-induced exacerbation (EX) vs 7-14 days post-exacerbation (post-EX). B. Normalized count levels comparing type 2 high vs type 2 low nasal brushings for genes that showed IGF2BP3-dependent IL-13-modulation in our Frac-seq data . C: Heatmap showing the normalized expression values of the common mRNA signature between differentially expressed genes in the GALA II cohort and our Frac-seq IGF2BP3 and IL-13-dependent DEGs. D. Model for IGF2BP3 effects on type 2 responses. Decreasing IGF2BP3 levels leads to increased stability of IL13RA1 and IL4R mRNA and increased surface expression of IL4R and IL13Rα1, leading to increased STAT6 phosphorylation and increased IL-13 (and possibly IL-4) transcriptional responses, contributing to an overall increased type 2 response. In disease, IGF2BP3 is upregulated potentially to supress excessive T2 responses.

Article Snippet: Primary bronchial epithelial cells (BECs) were commercially sourced (PromoCell), grown and expanded in growth factor-supplemented medium (PromoCell) on collagen-coated plates.

Techniques: Expressing, Control, Microarray, Phospho-proteomics

Journal: bioRxiv

Article Title: The RNA binding protein Insulin Growth factor 2 binding Protein 3 (IGF2BP3) modulates IL-13/IL-4 signalling in human bronchial epithelial cells and is dysregulated in type 2 disease

doi: 10.1101/2025.07.19.665669

Figure Lengend Snippet: Decreasing IGF2BP3 levels increases IL-13/IL-4-driven transcriptional responses. RT-qPCR results of primary BECs transfected with siRNAs against IGF2BP3 (siIGF2BP3) or scrambled control (siControl) and stimulated with IL-13 or IL-4. A: RT-qPCR results for IGF2BP3 knock down and the major signalling mediators of IL-13 and IL-4. B: RT-qPCR results for chemokines. C: RT-qPCR results for pro-inflammatory IL6 and IL8 mRNAs. Data (n=3) depicted as mean ±SEM. One-tailed ratio t-tests. * P ≤ 0.05; ** P ≤ 0.01.

Article Snippet: Primary bronchial epithelial cells (BECs) were commercially sourced (PromoCell), grown and expanded in growth factor-supplemented medium (PromoCell) on collagen-coated plates.

Techniques: Quantitative RT-PCR, Transfection, Control, Knockdown, One-tailed Test