1. Advanced co-culture 3D breast cancer model for investigation of fibrosis induced by external stimuli: optimization study
- Author
-
Aurélie François, Lina Bezdetnaya, Ilya Yakavets, Alice Benoit, Jean-Louis Merlin, Guillaume Vogin, Centre de Recherche en Automatique de Nancy (CRAN), Université de Lorraine (UL)-Centre National de la Recherche Scientifique (CNRS), Institut de Cancérologie de Lorraine - Alexis Vautrin [Nancy] (UNICANCER/ICL), UNICANCER, Ingénierie Moléculaire et Physiopathologie Articulaire (IMoPA), and Centre François Baclesse, Centre National de Radiothérapie du Grand Duché du Luxembourg
- Subjects
Cellular homeostasis ,Breast Neoplasms ,[SDV.CAN]Life Sciences [q-bio]/Cancer ,Models, Biological ,Article ,03 medical and health sciences ,0302 clinical medicine ,Breast cancer ,Fibrosis ,Spheroids, Cellular ,medicine ,Tumor Cells, Cultured ,Humans ,Cancer models ,030304 developmental biology ,0303 health sciences ,Tumor microenvironment ,Multidisciplinary ,Radiotherapy ,Chemistry ,Spheroid ,medicine.disease ,Coculture Techniques ,3. Good health ,030220 oncology & carcinogenesis ,Cancer cell ,Cancer research ,MCF-7 Cells ,Immunohistochemistry ,Transforming growth factor - Abstract
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.
- Published
- 2020
- Full Text
- View/download PDF