Abstract LB-318: Automated monitoring and analysis of kinetic live cell 3D spheroid-based tumor invasion within a hydrogel matrix

Abstract

Abstract Metastasis is the main cause of death in cancer patients and one of the most complex biological processes in human diseases. The development of therapies designed to forestall the metastatic activity of tumors has been met with multiple challenges. The first is the initial focus on single target remedies. As many types of cancers develop multiple mutations during tumor progression, individual cancers are often little affected by this type of drug. The advancement of novel methods that allow for the discernment of the effect a potential therapy has on the invasive phenotype of a particular type of cancer has proven invaluable to circumventing these early failures. A second hurdle is the choice of an appropriate cell model. Tumors in vivo exist as a three-dimensional (3D) mass of multiple cell types, including cancer and stromal cells. Therefore, incorporating a 3D spheroid-type cellular structure that includes co-cultured cell types forming a tumoroid, provides a more predictive model than the use of individual cancer cells cultured on the bottom of a well in traditional two-dimensional (2D) format. A third impediment is the inclusion of proper incubation time. Traditionally in vitro assessments of anti-cancer drugs have included short-term incubations, sometimes limited to 48 hours. In vivo treatments, however, can consist of a multi-week regimen. Pre-clinical tests must also, then, extend a similar length of time. A final hindrance which is necessary to overcome is the proper capture and analysis of microscopic images. The ability to monitor the entire tumoroid with a degree of sensitivity across multiple z-planes as cells invade into the matrix, as well as quantify changes over time, is critical. The information presented here will demonstrate a combined procedure for the generation of 3D spheroidal tumoroid structures, creation of a suitable invasion matrix, automated kinetic image-based monitoring of tumor invasion, and cellular analysis of captured z-stacked images. Citation Format: Brad Larson. Automated monitoring and analysis of kinetic live cell 3D spheroid-based tumor invasion within a hydrogel matrix [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2018; 2018 Apr 14-18; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2018;78(13 Suppl):Abstract nr LB-318.