Accurate Identification of Cancer Cells in Complex Pre‐Clinical Models Using a Deep‐Learning Neural Network: A Transfection‐Free Approach

3D co-cultures are key tools for in vitro biomedical research as they recapitulate more closely the in vivo environment while allowing a tighter control on the culture's composition and experimental conditions. The limited technologies available for the analysis of these models, however, hamper their widespread application. The separation of the contribution of the different cell types, in particular, is a fundamental challenge. In this work, ORACLE (OvaRiAn Cancer ceLl rEcognition) is presented, a deep neural network trained to distinguish between ovarian cancer and healthy cells based on the shape of their nucleus. The extensive validation that are conducted includes multiple cell lines and patient-derived cultures to characterize the effect of all the major potential confounding factors. High accuracy and reliability are maintained throughout the analysis (F1_score> 0.9 and Area under the ROC curve -ROC-AUC- score = 0.99) demonstrating ORACLE's effectiveness with this detection and classification task. ORACLE is freely available (https://github.com/MarilisaCortesi/ORACLE/tree/main) and can be used to recognize both ovarian cancer cell lines and primary patient-derived cells. This feature is unique to ORACLE and thus enables for the first time the analysis of in vitro co-cultures comprised solely of patient-derived cells.