Culturing Human Lung Adenocarcinoma Cells in a Serum-Free Environment

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.

Keywords: A549; Air–liquid interface; Cell culture; Cell lines; Chemically defined media; Fetal bovine serum; Serum-free media; Xeno-free media.