Development of Apical-out Airway Organoids to Evaluate Respiratory Toxicity of Polystyrene Microplastics
Bibliographic record
Abstract
Background: Even though it has been reported that microplastic could infiltrate in the lung and induce inflammation, exact pathophysiology of microplastic indued-lung injuries has not been fully revealed.One of the main hurdles for evaluating respiratory exposure effect of microplastic on the airway is shortage of in vitro models which are relevant with human exposure of microplastic during inhalation.In this study, we generated apical out airway organoid (AOAO) which epithelial cell located in outer surface of organoid to evaluate the feasibility of AOAO for respiratory exposure models of microplastics.Methods: Human airway basal stem cells (hABSCs, passage 2) were trypsinized and resuspended (10,000 cells/mL) in Pneuma Cult-ALI Medium (#05001; STEMCELL Technologies, Canada) supplement with Pneuma Cult-ALI supplements, hydrocortisone, and heparin.One hundred µL of resuspended hABSCs were placed per well in a low attachment 96-well microplate (Corning #7007, USA) and centrifuged at 1,500 RPM for 15 minutes.Fifty µL of medium were removed and refreshed every 2 days.The cultures were maintained at 37 °C with 5% CO2 for 21 days, and generation and movement of AOAOs were video recorded at 7-day intervals using an EVOS M7000 Imaging System (ThermoFisher Scientific, USA).After generating AOAO, polystyrene (PS) was treated and inflammation response were evaluated.Results: To characterize AOAO in the resulting day 21, we performed immunofluorescence staining of key markers of airway epithelium and observed highly selective localization of acetylated-alpha-tubulin on the organoid outer surface.Besides ciliated cells, other cell types of airway epithelium such as basal cells and goblet cells were identified.Moreover, goblet cells and basal cells were localized on the inner layer of AOAOs.We then calculated the length of cilia on the organoid's outer surface.Similar to the human airway tissue, AOAOs have beating cilia.By treating PS, goblet cell hyperplasia was shown in the AOAO.Expression of inflammatory cytokine such as tumor necrosis factor-α and interleukin-1β were increased by PS.Conclusion: Apical-out airway organoids mimic human upper airway epithelium and cilia function.Moreover, AOAOs represent an increase of mucin secretion and inflammation following microplastic exposure.
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How this classification was reachedexpand
Full frame machine prediction
Teacher imitationNot calibrated prevalence, not ground truth. Human validation pending. The Gemma side is a direct model label for every work in the frame, read from the title-only record. The Codex side is a classifier learned from the 10,348 direct Codex labels and calibrated to design-weighted sample rates; fields without enough sample support carry no Codex call. Candidate is the union of the two sides; consensus is their intersection. These outputs are machine_predicted_unvalidated and are not human labels.
Distilled classifier scores by category (both heads)
| Category | Codex | Gemma |
|---|---|---|
| Metaresearch | 0.001 | 0.000 |
| Meta-epidemiology (narrow) | 0.001 | 0.000 |
| Meta-epidemiology (broad) | 0.000 | 0.001 |
| Bibliometrics | 0.000 | 0.000 |
| Science and technology studies | 0.000 | 0.000 |
| Scholarly communication | 0.000 | 0.000 |
| Open science | 0.000 | 0.000 |
| Research integrity | 0.001 | 0.001 |
| Insufficient payload (model declined to judge) | 0.001 | 0.001 |
Machine scores (provisional)
The two teacher heads of the student model, read on this work. A score orders the frame for review; it never asserts a category, and the validation status ships verbatim with every row.
Baseline scores from an immature model (maturity gate not passed, 7 training rounds). Scores rank; they never assert a category.
score_only:v0-immature-baseline · verbatim from the scoring run: score_only means the number may rank works, and no category label ships from itClassification
machine, unvalidatedMachine predicted; a candidate call from one source (direct Gemma or distilled Codex), not a consensus.
How this classification was reached, model by model and score by score, is at the end of the page under "How this classification was reached".