Advanced co-culture 3D breast cancer model for investigation of fibrosis induced by external stimuli: optimization study.
Fiche publication
Date publication
décembre 2020
Journal
Scientific reports
Auteurs
Membres identifiés du Cancéropôle Est :
Dr BEZDETNAYA-BOLOTINE Lina, Pr MERLIN Jean-Louis
Tous les auteurs :
Yakavets I, Francois A, Benoit A, Merlin JL, Bezdetnaya L, Vogin G
Lien Pubmed
Résumé
Radiation-induced fibrosis (RIF) is the main late radiation toxicity in breast cancer patients. Most of the current 3D in vitro breast cancer models are composed by cancer cells only and are unable to reproduce the complex cellular homeostasis within the tumor microenvironment to study RIF mechanisms. In order to account complex cellular interactions within the tumor microenvironment, an advanced 3D spheroid model, consisting of the luminal breast cancer MCF-7 cells and MRC-5 fibroblasts, was developed. The spheroids were generated using the liquid overlay technique in culture media into 96-well plates previously coated with 1% agarose (m/v, in water). In total, 21 experimental setups were tested during the optimization of the model. The generated spheroids were characterized using fluorescence imaging, immunohistology and immunohistochemistry. The expression of ECM components was confirmed in co-culture spheroids. Using α-SMA staining, we confirmed the differentiation of healthy fibroblasts into myofibroblasts upon the co-culturing with cancer cells. The induction of fibrosis was studied in spheroids treated 24 h with 10 ng/mL TGF-β and/or 2 Gy irradiation. Overall, the developed advanced 3D stroma-rich in vitro model of breast cancer provides a possibility to study fibrosis mechanisms taking into account 3D arrangement of the complex tumor microenvironment.
Référence
Sci Rep. 2020 Dec 4;10(1):21273