Here, we investigated the heterogeneity of the 16HBE14o− cell line, particularly with respect to CFTR/CFTR expression since this line and its gene‐edited derivatives are frequently used as the model system of choice in CF research (Bednarski et al., 2016; Erwood et al., 2020; Ko et al., 2022; Michaels et al., 2022; Santos et al., 2022, 2023; Valley et al., 2019). Here, CFTR is linked to cystic fibrosis.