Inter-laboratory variability of A549 epithelial cells grown under submerged and air-liquid interface conditions
Language English Country England, Great Britain Media print-electronic
Document type Journal Article
PubMed
33905840
DOI
10.1016/j.tiv.2021.105178
PII: S0887-2333(21)00103-X
Knihovny.cz E-resources
- Keywords
- Alveolar epithelial cells, In vitro model, Inter-laboratory assessment, Lung model, Standard operating procedure,
- MeSH
- Cell Differentiation drug effects MeSH
- Cell Culture Techniques * MeSH
- A549 Cells MeSH
- Epithelial Cells * drug effects MeSH
- Interleukin-6 metabolism MeSH
- Interleukin-8 metabolism MeSH
- Laboratories MeSH
- Humans MeSH
- Lipopolysaccharides pharmacology MeSH
- Cell Proliferation drug effects MeSH
- Reproducibility of Results MeSH
- Tumor Necrosis Factor-alpha pharmacology MeSH
- Cell Survival drug effects MeSH
- Check Tag
- Humans MeSH
- Publication type
- Journal Article MeSH
- Names of Substances
- CXCL8 protein, human MeSH Browser
- IL6 protein, human MeSH Browser
- Interleukin-6 MeSH
- Interleukin-8 MeSH
- Lipopolysaccharides MeSH
- Tumor Necrosis Factor-alpha MeSH
In vitro cell models offer a unique opportunity for conducting toxicology research, and the human lung adenocarcinoma cell line A549 is commonly used for toxicology testing strategies. It is essential to determine whether the response of these cells grown in different laboratories is consistent. In this study, A549 cells were grown under both submerged and air-liquid interface (ALI) conditions following an identical cell seeding protocol in two independent laboratories. The cells were switched to the ALI after four days of submerged growth, and their behaviour was compared to submerged conditions. The membrane integrity, cell viability, morphology, and (pro-)inflammatory response upon positive control stimuli were assessed at days 3, 5, and 7 under submerged conditions and at days 5, 7, and 10 at the ALI. Due to the high variability of the results between the two laboratories, the experiment was subsequently repeated using identical reagents at one specific time point and condition (day 5 at the ALI). Despite some variability, the results were more comparable, proving that the original protocol necessitated improvements. In conclusion, the use of detailed protocols and consumables from the same providers, special training of personnel for cell handling, and endpoint analysis are critical to obtain reproducible results across independent laboratories.
References provided by Crossref.org
Computational Design of Pore-Forming Peptides with Potent Antimicrobial and Anticancer Activities
The Effect of Chronic Exposure of Graphene Nanoplates on the Viability and Motility of A549 Cells
