Journal: Environmental Analysis, Health and Toxicology

Article Title: Development of a multi-analysis model using an epithelial-fibroblast co-culture system as an alternative to animal testing

doi: 10.5620/eaht.2024024

Figure Lengend Snippet: Differentiation of Normal Human Bronchial Epithelial Cells under ALI : After cultured hAECB cells in PneumaCultTM-ALI media for 4 weeks at ALI conditions, differentiated pseudostratified epithelial layer was confirmed. (a) The TEER value measured under ALI conditions maintained above 400 Ω.cm 2 for 4 weeks. (b) Hematoxylin & eosin (H&E) staining of hAECB cells differentiated for over 4 weeks. (c) Immunofluorescence staining of the nuclear (DAPI, blue), goblet cells (MUC5AC, green) of hAECB cells differentiated for over 4 weeks.

Article Snippet: The primary human airway epithelial cells from bronchi (hAECB), isolated from human lung biopsies, were purchased from Epithelix Sarl (Swizerland).

Techniques: Cell Culture, Staining, Immunofluorescence

Journal: Environmental Analysis, Health and Toxicology

Article Title: Development of a multi-analysis model using an epithelial-fibroblast co-culture system as an alternative to animal testing

doi: 10.5620/eaht.2024024

Figure Lengend Snippet: Functionality analysis using 3D Epithelial-fibroblast co-culture system after 3 h of exposure to TGF-β1. (a) Differentiated hAECB cells were stained with hematoxylin & eosin (H&E), and changes in the cilia were observed after exposure to TGF-β1. (b) The cilia beating area of differentiated epithelial cells under ALI conditions. The cilia beating area was visualized by distinguishing between an area where beating was not detected and an area where beating was detected. The cilia beating area is the percentage (%) of the area with movement relative to all pixels in the FOV, calculated as the mean and median (25th percentile to 75th percentile). (c) To observe MRC-5 cell migration, the cells were co-cultured with hAECB cells treated with TGF-β1 for 9 h. Cell migration was observed microscopically over time and expressed relative to control (fold induction). (d) Conditioned medium obtained from the basal region was collected and exposed to a collagen gel seeded with MRC-5 cells. Collagen contraction was observed over time using a microscope and expressed compared to control (magnification-derived). Values are significantly different from control at *p < 0.05, **p < 0.01.

Article Snippet: The primary human airway epithelial cells from bronchi (hAECB), isolated from human lung biopsies, were purchased from Epithelix Sarl (Swizerland).

Techniques: Co-Culture Assay, Staining, Migration, Cell Culture, Control, Microscopy, Derivative Assay