haec medium  (Epithelix)

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Modeling airway persistent infection of Moraxella catarrhalis and nontypeable Haemophilus influenzae by using human in vitro models

doi: 10.3389/fcimb.2024.1397940

Figure Lengend Snippet: Characterization of ALI respiratory models. (A) LSCM analysis of epithelial differentiation: cilia (β-tubulin IV), mucus (MUC5AC), club cells (Uteroglobin), basal cells (p63) and Zonula Occludens 1 (ZO-1) were present in both standard and inverted models (white bars = 50µm). (B) Trans-epithelial electrical resistance measurement during epithelial differentiation. Results are expressed as mean values plus standard deviation calculated on 25 replicates for each configuration. (C) Evaluation of ciliated and goblet cells relative proportion based on the quantification of the fluorescent area related to β-tubulin IV and MUC5AC, respectively. Bars represent mean values plus standard deviation calculated on three replicates from three independent experiments (144 fields were analyzed for each sample).

Article Snippet: Cells were expanded in 75 cm 2 flasks by using human Airway Epithelial Cell Culture medium (hAEC; Epithelix) supplemented with Primocin (Invivogen), at 37°C in 5% CO 2 until approximately 80% confluence and used between passage 4 and 6.

Techniques: Standard Deviation

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Modeling airway persistent infection of Moraxella catarrhalis and nontypeable Haemophilus influenzae by using human in vitro models

doi: 10.3389/fcimb.2024.1397940

Figure Lengend Snippet: LSCM analysis of airway models infected with Mcat AERIS 415. (A) Mcat infection expands intraepithelially after 48 hours from the challenge (red is Mcat; grey is cell membrane; white bar = 5 µm). (B) Mcat colonies emerge apically from the epithelium. Only bacteria residing on the external perimeter of the aggregate (white arrows) can be efficiently immuno-stained with anti-UspA2 antibodies; white bar = 50 µm. (C) At seven days post infection, Mcat macroaggregates pervade the epithelial surface and most cells contain IBCs; the upper panel represent a Z/Y section of the epithelium while the lower panel is a X/Y view of the same stack (blue is DNA; yellow is F-actin; green are cilia; red is Mcat; white bar = 50 µm). (D, E) Scanning Electron Microscopy analysis of inverted airway models 14 days after Mcat inoculum shows the presence of an extra-polymeric substance enveloping bacterial clusters (white bar is 10 µm and 2 µm for (D, E) respectively). (F) Immunogold labeling assay conducted on Mcat biofilm ultrathin sections shows that UspA2 is expressed by bacterial cells residing inside the aggregate; white bar = 1 µm (G) dsDNA immunogold staining of a biofilm section; white bar = 0.5 µm.

Article Snippet: Cells were expanded in 75 cm 2 flasks by using human Airway Epithelial Cell Culture medium (hAEC; Epithelix) supplemented with Primocin (Invivogen), at 37°C in 5% CO 2 until approximately 80% confluence and used between passage 4 and 6.

Techniques: Infection, Membrane, Bacteria, Staining, Electron Microscopy, Labeling

Journal: Frontiers in Cellular and Infection Microbiology

Article Title: Modeling airway persistent infection of Moraxella catarrhalis and nontypeable Haemophilus influenzae by using human in vitro models

doi: 10.3389/fcimb.2024.1397940

Figure Lengend Snippet: LSCM and TEM analysis of airway models infected with NTHi Fi176. (A) After 72 hours of infection with strain Fi176 bacteria can be found adhering to the epithelial surface, entering or inside cells; white bar = 50 µm. (B) NTHi forms IBCs after 1 week of infection (red arrows). Bacteria residing in the internal part of IBCs are negative for an anti-Fi176 antibody unlike those located in the margin (stained in red and indicated by white arrowheads); white bar = 10 µm. (C) TEM analysis of IBCs residing below the apical surface of the epithelium (black arrows). Bacteria localized paracellularly are indicated by black arrowheads. Cell-cell junctions appear loose (black asterisks); white bar = 5 µm. (D) Higher magnification of an IBC formed by NTHi; white bar = 2 µm.

Article Snippet: Cells were expanded in 75 cm 2 flasks by using human Airway Epithelial Cell Culture medium (hAEC; Epithelix) supplemented with Primocin (Invivogen), at 37°C in 5% CO 2 until approximately 80% confluence and used between passage 4 and 6.

Techniques: Infection, Bacteria, Staining