Methodology for application of normal and malignant breast tissue-derived organoid culture as a model for diagnostic extracellular vesicle discovery

Development of new diagnostic approaches heavily relies on the experimental cancer models, including cell culture in vitro. Extracellular vesicles (EVs) recently have gained popularity as potential diagnostic biomarkers for various diseases including cancer. The aim of this project is to test different EV isolation methods from 2D and 3D breast cancer cell lines and breast cancer (BC) patient-derived organoids and blood for further application in identification of promising non invasive biomarkers for BC diagnostics and theranostics. During the project a sets of biological analytical samples (EVs, RNA from mentioned material) will be obtained. The project involves isolation, characterization and analysis of EVs and their microRNA cargo. This pilot project is the first step toward further project applications that will allow to explore the potential of organoid culture system and to determine diagnostic marker candidates for further analysis and validation in a larger population.

In this project we are testing several extracellular vesicle isolation methods to further choose the best for isolation of EVs from breast cancer patient-derived organoids and blood. This pilot project is the first step towards identification of different molecules in blood that could be derived from breast normal not malignant tissues.