Journal: Biomaterials Research

Article Title: Advanced pathophysiology mimicking lung models for accelerated drug discovery

doi: 10.1186/s40824-023-00366-x

Figure Lengend Snippet: Morphological appearance, electrical resistance, and FITC-Dextran permeability of the optimised models (static 10,000 BEAS-2B cells, static 10,000 BEAS2B-A549, CNBio BEAS2B-A549 and CNBio 10,000 A549-BEAS2B). A The morphology of cells at the end of the culture for all the models on Day 11. Scale bar represents 200 µm; B The electrical resistance of all the models over a period of 11 days where ALI was performed on Day 4; C The permeability of FITC-Dextran into the basolateral compartment of all the models. Negative (-ve) control was the medium only without FITC-Dextran. Positive (+ ve) control was empty insert without cells followed the same protocol as the inserts with cells. Mean ± SD are plotted. Statistical significance was tested using ordinary one-way ANOVA, followed by Dunn’s multiple comparison’s test for pair-wise comparisons, n = 3. D Representative IF staining of CNBio 10,000 BEAS2B-A549 and CNBio 10,000 A549-BEAS2B showing disrupted tight junctional proteins ZO-1 (red) and DAPI (blue); scale bar, 50 µm

Article Snippet: PneumaCultTM-ALI medium (STEMCELL TechnologiesTM, Australia) was used during the last three weeks of differentiation of the models in air liquid interface (ALI) condition (PneumaCultTM-ALI medium was used in basolateral compartment and no medium was used in the apical compartment).

Techniques: Permeability, Control, Staining

Journal: Biomaterials Research

Article Title: Advanced pathophysiology mimicking lung models for accelerated drug discovery

doi: 10.1186/s40824-023-00366-x

Figure Lengend Snippet: Comparison of morphological appearance, barrier integrity, permeability and structure of developed models using human primary cell lines. Representative morphological appearance, SEM images and IF staining of developed models; A static mono-cell culture (NHBE), B static co-culture (NHBE-NHLF), C dynamic mono-cell culture (CNBio NHBE), D dynamic co-culture (CNBio NHBE-NHLF) on the Day 25. The morphology of cells at the end of culture (Day 25 of culture). Scale bar represents 200 µm. For SEM, images showed the morphology of cells on the apical surface of the model; scale bar, 10 µm. For IF staining, ZO-1 (red) and DAPI (blue) staining were used to demonstrate the continuous tight junctional connections; scale bar, 50 µm. E The electrical resistance of the models over a period of 25 days of culture; ALI condition was performed on Day 7 for all models; F The permeability of FITC-Dextran into the basolateral compartment of all the models. Negative (-ve) control was the medium only without FITC-Dextran. Positive (+ ve) control was the empty insert without cells followed the same protocol as the inserts with cells. Error bars denote standard deviation. Statistical significance was shown between positive (+ ve) control and CNBio NHBE; **: P < 0.01. G Representative histological cross-sections (H&E staining) and IF staining display essential epithelial features, including acetylated α-tubulin (green, marker for ciliated cells) and MUC5B (red, marker for goblet cells) for the static culture and dynamic culture. Objective 40x, scale bar represents 50 µm

Article Snippet: PneumaCultTM-ALI medium (STEMCELL TechnologiesTM, Australia) was used during the last three weeks of differentiation of the models in air liquid interface (ALI) condition (PneumaCultTM-ALI medium was used in basolateral compartment and no medium was used in the apical compartment).

Techniques: Comparison, Permeability, Staining, Cell Culture, Co-Culture Assay, Control, Standard Deviation, Marker