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Inter-laboratory variability of A549 epithelial cells grown under submerged and air-liquid interface conditions
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SYSNO ASEP 0552840 Document Type J - Journal Article R&D Document Type Journal Article Subsidiary J Článek ve WOS Title Inter-laboratory variability of A549 epithelial cells grown under submerged and air-liquid interface conditions Author(s) Bartošová, Hana (UEM-P)
Meldrum, K. (GB)
Karakocak, B.B. (CH)
Balog, S. (CH)
Doak, S.H. (GB)
Petri-Fink, A. (CH)
Clift, M.J.D. (US)
Rothen-Rutishauser, B. (CH)Article number 105178 Source Title Toxicology in Vitro. - : Elsevier - ISSN 0887-2333
Roč. 75, sep. (2021)Number of pages 10 s. Language eng - English Country GB - United Kingdom Keywords standard operating procedure ; in vitro model ; alveolar epithelial cells ; lung model ; inter-laboratory assessment OECD category Toxicology Method of publishing Open access Institutional support UEM-P - RVO:68378041 UT WOS 000672807200021 EID SCOPUS 85105355880 DOI 10.1016/j.tiv.2021.105178 Annotation 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. Workplace Institute of Experimental Medicine Contact Lenka Koželská, lenka.kozelska@iem.cas.cz, Tel.: 241 062 218, 296 442 218 Year of Publishing 2022 Electronic address https://www.sciencedirect.com/science/article/pii/S088723332100103X?via%3Dihub
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