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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