bronchial epithelial cell growth medium  (PromoCell)

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: SARS-CoV-2 pseudovirus ex vivo infected nasal epithelial cells (n=3 pools). (E-H) SARS-CoV-2 pseudovirus ex vivo infected lung epithelial cells (n= 9). (I-L) SARS-CoV-2 pseudovirus ex vivo infected renal cortex cells (n=7). Flow cytometry plots showing the phenotypic comparison between GFP⁺ (infected, green) and GFP⁻ (uninfected) cells of CD45 - CD31 - EpCAM + cells from the (A) pool #NAL02 (n= 7) or (E) #HLTE197, which includes a paired sample exposed to a spike-empty pseudovirus (background). (B and F) Violin plots depicting the frequency (%) of different epithelial marker expressions within total nasal ( B ) or pulmonary ( F ) EpCAM + (grey) and EpCAM + GFP + cells (green). (C-D and G-H) Boolean pie charts displaying the proportion of nasal ( C-D ) or pulmonary ( G-H ) EpCAM + ( C-G ) and EpCAM + GFP + ( D-H ) cells expressing combinations of color code molecules according to the legend and indicated as surrounding arcs around the pie chart. (I) Flow cytometry plots showing the phenotypic comparison of CD45 - CD31 - cells from #RINN22, either infected (GFP⁺; green) or exposed to a ‘background’ pseudovirus. (J) Violin plots depicting the frequency (%) of various molecules within total CD31 - (grey) and CD31 - GFP + cells (green). (K-L) Boolean pie charts displaying the proportion of CD31 - ( K ) and CD31 - GFP + ( L ) cells expressing combinations of color code molecules according to the legend and indicated as surrounding arcs around the pie chart. For all violin plots, data are represented as median ± IQR. Statistical analyses were performed using two-sided nonparametric Wilcoxon matched-pairs signed-rank test.

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Ex Vivo, Infection, Flow Cytometry, Comparison, Marker, Expressing

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: (A) UMAP projection of high-dimension single-cell flow cytometry nasal data of EpCAM + cells from ex vivo SARS-CoV-2 pseudovirus infected samples, depicting the differentiation of 8 clusters (MC) and relative abundance of each cluster across the three pools of nasal samples. (B) UMAP visualization highlighting the EpCAM + GFP + infected population (green). Adjacent heatmap displays normalized mean fluorescence intensity of epithelial markers across the 8 identified clusters as indicated in (A) . (C) UMAP-based visualization of the spatial expression of six molecules (top) and density histograms comparing EpCAM⁺ and EpCAM⁺GFP⁺ populations (bottom).

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Single Cell, Flow Cytometry, Ex Vivo, Infection, Fluorescence, Expressing

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: (A) UMAP projection of high-dimension single-cell flow cytometry data of pulmonary EpCAM + cells from uninfected or ex vivo SARS-CoV-2 pseudovirus infected samples, depicting the differentiation of 8 clusters (MC) and relative abundance of each cluster across uninfected versus infected samples (bars on top). Bars below show the percentage of less represented clusters (MC03-MC08). (B) UMAP visualization highlighting the EpCAM + GFP + infected population (green). Adjacent heatmap (bottom) displays normalized mean fluorescence intensity of epithelial markers across the 8 identified clusters as indicated in (A) highlighting the cluster representing GFP + cells (MC07). (C) Volcano plot displaying the differential cluster abundance comparing uninfected and infected samples, the green dot corresponds to MC07. (D) UMAP-based visualization of the spatial expression of six molecules (top) and density histograms comparing EpCAM⁺ and EpCAM⁺GFP⁺ populations (bottom).

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Single Cell, Flow Cytometry, Ex Vivo, Infection, Fluorescence, Expressing

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: (A) UMAP projection of high-dimension single-cell flow cytometry data of renal CD31 - cells from uninfected or ex vivo SARS-CoV-2 pseudovirus infected samples, depicting the differentiation of 17 clusters (MC) and relative abundance of each cluster across uninfected versus infected samples. (B) UMAP visualization highlighting the CD31 - GFP + infected population (green). Adjacent heatmap displays normalized mean fluorescence intensity of epithelial markers across the 17 identified clusters as indicated in ( A ) highlighting the cluster representing GFP + cells (MC05). (C) Volcano plot displaying the differential cluster abundance comparing uninfected and infected samples, the green dot corresponds to MC05. (D) UMAP-based visualization of the spatial expression of six molecules (top) and density histograms comparing CD31 - and CD31 - GFP + populations (bottom).

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Single Cell, Flow Cytometry, Ex Vivo, Infection, Fluorescence, Expressing

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: (A) Schematic overview of the experimental workflow. Epithelial cells were isolated from nasal, lung, and kidney tissues and infected ex vivo with SARS-CoV-2 pseudovirus. GFP⁺ (infected) and GFP⁻ (uninfected) cells were sorted and processed using Smart-seq2. After quality control, cells were clustered and analyzed by tissue. (B–D) Nasal epithelial cells. (B) UMAP embedding of five transcriptionally distinct clusters. (C) Cluster distribution across all nasal epithelial cells. (D) Cluster proportions stratified by GFP⁺ and GFP⁻ conditions. (E–G) Lung epithelial cells. (E) UMAP embedding of lung-derived cells showing the separation of three epithelial and one stromal population. (F) Cluster distribution across epithelial and stromal populations. (G) Cluster proportions by infection status. (H–J) Renal cortex epithelial cells. (H) UMAP embedding of two epithelial clusters. (I) Cluster distribution across all renal epithelial cells. (J) Cluster proportions stratified by GFP⁺ and GFP⁻ conditions.

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Isolation, Infection, Ex Vivo, Control, Derivative Assay

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: (A) Violin plots showing distributions of standard single-cell RNA-seq quality control metrics across all cells, including the number of detected genes (nFeature_RNA), total UMI counts (nCount_RNA), and the percentage of mitochondrial transcripts (percent.mt). ( B ) Heatmap of the top 10 differentially expressed genes for each identified cluster. Genes are ranked by average log-normalized expression within each cluster relative to others. Color scale indicates scaled expression levels. ( C ) Violin plots depicting normalized expression levels of selected epithelial marker genes ( EPCAM, ELF3, CLDN4 , and CDH1 ) across the indicated clusters, highlighting cluster-specific expression patterns. ( D ) Dot plot summarizing the expression of representative marker genes across clusters, based on Ahn, J.H., et al. (2021. J. Clin. Invest). Dot size reflects the percentage of cells expressing each gene, and color intensity represents the average expression level within each cluster

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Single Cell, RNA Sequencing, Control, Expressing, Marker

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: (A) Violin plots showing distributions of standard single-cell RNA-seq quality control metrics across all cells, including the number of detected genes (nFeature_RNA), total UMI counts (nCount_RNA), and the percentage of mitochondrial transcripts (percent.mt). (B) Heatmap of the top 10 differentially expressed genes per cluster, highlighting transcriptionally distinct epithelial and stromal compartments. Genes are ranked by average log-normalized expression within each cluster relative to others. Color scale indicates scaled expression levels. (C) Violin plots depicting normalized expression levels of representative markers: epithelial markers EPCAM and ELF3 enriched in epithelial groups, and stromal/extracellular matrix markers DCN and MFAP4 enriched in fibroblasts.

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Single Cell, RNA Sequencing, Control, Expressing

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: (A) Violin plots showing distributions of standard single-cell RNA-seq quality control metrics across all cells, including the number of detected genes (nFeature_RNA), total UMI counts (nCount_RNA), and the percentage of mitochondrial transcripts (percent.mt). (B) Heatmap of the top 10 differentially expressed genes per cluster, separating proximal tubular epithelial cells from broader epithelial populations. Genes are ranked by average log-normalized expression within each cluster relative to others. Color scale indicates scaled expression levels. (C) Violin plots depicting normalized expression levels of representative markers: epithelial marker EPCAM enriched in epithelial cells, proximal-tubule markers AQP1 and CUBN enriched in the proximal tubular epithelial cluster and SLC12A3 , expressed in the distal convoluted tubule.

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Single Cell, RNA Sequencing, Control, Expressing, Marker

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: Volcano plots showing differentially expressed genes between GFP⁺ and GFP⁻ in overall epithelial cells from the (A) nasal mucosa, (B) lung parenchyma and (C) renal cortex; or in individual clusters from a given tissue: (D) epithelial cluster 0 from the nasal mucosa; (F) alveolar type 2 (AT2) cells from the lung; and (G) general epithelial cluster from renal cortex. The horizontal axis shows log₂ fold change, and the vertical axis shows –log₁₀ adjusted p values. Selected significantly upregulated genes (in red) are highlighted in bigger dots.

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques:

Journal: bioRxiv

Article Title: Site-Specific Entry Factors Define Cellular Susceptibility to SARS-CoV-2 in Human Tissues

doi: 10.64898/2026.05.07.723425

Figure Lengend Snippet: ( A–B ) SARS-CoV-2 pseudovirus entry in lung ( A ) and kidney ( B ) epithelial cells following treatment with inhibitors. Lung or renal cortex-derived cells were exposed to pseudovirus in the presence of anti-ACE2 (25 µg/ml), Camostat (100 µM), KP-457 (ADAM17 inhibitor, 100 µM), anti-IL1R1 (100 µM), Ruxolitinib (JAK1/2 inhibitor, 100 µM), anti-ADAMTSL3 (250 ng/ml), anti-CADM1 (625 ng/ml), anti-GULP1 (625 ng/ml), anti-MDGA2 (62.5 ng/ml), anti-PILRα (1.25 µg/ml) or anti-PTPRK (1.25 µg/ml). Infection levels, quantified by luciferase activity, are expressed relative to untreated controls (100% infection). Each color-coded dot indicates an individual tissue with median and interquartile range indicated for each treatment with dotted lines. Statistical significance was assessed using a Kruskal–Wallis test for multiple comparisons (****P < 0.0001, ***P < 0.001, **P < 0.01, *P < 0.05). ( C ) Plots showing Spearman correlation coefficients and associated significant P values comparing inhibitor effects across lung and kidney tissues.

Article Snippet: Nasal epithelial cells were obtained with an ASI Rhino-Pro® nasal curette (Arlington, IL, USA) into Bronchial Epithelial Cell Medium (BEpiCM) (Innoprot) washed with PBS with 5 mM EDTA, incubated on a shaker (15 min. 30 rpm, 4°C) centrifuged, filtered and counted.

Techniques: Derivative Assay, Infection, Luciferase, Activity Assay

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