Standardization of esophageal adenocarcinoma in vitro model and its applicability for model drug testingTratnjek, Larisa (Avtor) Sibinovska, Nadica (Avtor) Kralj, Slavko (Avtor) Makovec, Darko (Avtor) Kristan, Katja (Avtor) Erdani-Kreft, Mateja (Avtor) esophageal adenocarcinomaFLO-1 cell linedisease pathophysiologybiological techniquescancercell biologydiseasesdrug discoveryengineeringoncologyFLO-1 cell line represents an important tool in esophageal adenocarcinoma (EAC) research as a verifed and authentic cell line to study the disease pathophysiology and antitumor drug screenings. Since in vitro characteristics of cells depend on the microenvironment and culturing conditions, we performed a thorough characterization of the FLO-1 cell line under diferent culturing conditions with the aim of (1) examining the efect of serum-free growth medium and air–liquid interface (A–L) culturing, which better refect physiological conditions in vivo and (2) investigating the diferentiation potential of FLO-1 cells to mimic the properties of the in vivo esophageal epithelium. Our study shows that the composition of the media infuenced the morphological, ultrastructural and molecular characteristics of FLO-1 cells, such as the expression of junctional proteins. Importantly, FLO-1 cells formed spheres at the A–L interface, recapitulating key elements of tumors in the esophageal tube, i.e., direct contact with the gas phase and three-dimensional architecture. On the other hand, FLO-1 models exhibited high permeability to model drugs and zero permeability markers, and low transepithelial resistance, and therefore poorly mimicked normal esophageal epithelium. In conclusion, the identifed efect of culture conditions on the characteristics of FLO-1 cells should be considered for standardization, data reproducibility and validity of the in vitro EAC model. Moreover, the sphere-forming ability of FLO-1 cells at the A–L interface should be considered in EAC tumor biology and anticancer drug studies as a reliable and straightforward model with the potential to increase the predictive efciency of the current in vitro approaches.20212023-03-14 14:36:48Članek v reviji144815sl