Culturing Human Lung Adenocarcinoma Cells in a Serum-Free Environment (original) (raw)

Abstract

The human lung adenocarcinoma cell line A549 is commonly used in cancer research as a model of malignant alveolar type II epithelial cells. A549 cells are frequently cultured in Ham’s F12K (Kaighn’s) or Dulbecco’s Modified Eagle’s Medium (DMEM), supplemented with glutamine and 10% fetal bovine serum (FBS). However, the use of FBS presents significant scientific concerns, such as the presence of undefined components and batch-to-batch variation leading to possible reproducibility issues in experiments and readouts. This chapter describes how to transition A549 cells to FBS-free medium and gives some insights on the further characterizations and functionality assays that would be necessary to perform for the validation of the cultured cells.

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References

  1. Lieber M, Todaro G, Smith B, Szakal A, Nelson-Rees W (1976) A continuous tumor-cell line from a human lung carcinoma with properties of type II alveolar epithelial cells. Int J Cancer 17:62–70
    Article CAS PubMed Google Scholar
  2. Lacroix G et al (2018) Air–liquid interface in vitro models for respiratory toxicology research: consensus workshop and recommendations. Appl In Vitro Toxicol 4:91–106
    Article PubMed PubMed Central Google Scholar
  3. Hartung T et al (2004) A modular approach to the ECVAM principles on test validity. Alternat Lab Anim 32:467–472
    Article CAS Google Scholar
  4. Barosova H et al (2021) Inter-laboratory variability of A549 epithelial cells grown under submerged and air-liquid interface conditions. Toxicol in Vitro 75:105178
    Article CAS PubMed Google Scholar
  5. Chary A et al (2022) Maximizing the relevance and reproducibility of A549 cell culture using FBS-free media. Toxicol in Vitro 105423. https://doi.org/10.1016/j.tiv.2022.105423
  6. van der Valk J et al (2010) Optimization of chemically defined cell culture media – replacing fetal bovine serum in mammalian in vitro methods. Toxicol in Vitro 24:1053–1063
    Article PubMed Google Scholar
  7. van der Valk J (2022) Fetal bovine serum – a cell culture dilemma. Science. https://doi.org/10.1126/science.abm1317

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Acknowledgments

This work was in part supported by the LIST internal project VEGALI (Esch-sur-Alzette, Luxembourg) and PETA Science Consortium International e.V. (Stuttgart, Germany).

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Authors and Affiliations

  1. Environmental Research and Innovation (ERIN) Department, Luxembourg Institute of Science and Technology (LIST), Esch-sur-Alzette, Luxembourg
    Aline Chary

Corresponding author

Correspondence toAline Chary .

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Editors and Affiliations

  1. Trinity Translational Medicine Institute, Trinity Centre for Health Sciences, Dublin, Ireland
    Dania Movia
  2. Trinity Translational Medicine Institute, Trinity Centre for Health Sciences, Dublin, Ireland
    Adriele Prina-Mello

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Chary, A. (2023). Culturing Human Lung Adenocarcinoma Cells in a Serum-Free Environment. In: Movia, D., Prina-Mello, A. (eds) Cancer Cell Culture. Methods in Molecular Biology, vol 2645. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-3056-3\_8

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