Abstract 1431: An optimized in vitro method to evaluate primary tumor cell invasion using optical imaging technology

Abstract

Abstract The recent development of human-in-mouse tumor models has provided better representative working models to understand tumor initiation and progression than previous mouse tumor models or cell lines. However, it has not been feasible to use these models for high-throughput functional or therapeutic screening. Our aim was to establish a complementary in vitro culturing system for high-throughput functional studies or therapeutic screening of breast cancer stem cells (BCSCs). We hypothesize that with extracellular matrix and stromal cell support, three-dimensional (3D) cultures can support the survival and invasion of primary human tumors in vitro. We report here a new method to objectively evaluate and quantify invasion of primary breast cancer tumor cells using Transwells and bioluminescence imaging (BLI) technology. We optimized the adhesion and invasion conditions for these primary tumor cells. We also labeled cells using Luc2-EGFP or -tdTomato lentiviral vectors so that the signal of all invaded cells could be quantified using BLI or fluorescence imaging of the entire bottom chamber of the Transwell system. Our results demonstrate that stromal cells are not only necessary for the survival of the primary tumor cells but also facilitate their invasion in vitro. This work establishes a new method for screening candidate genes, miRNAs and compounds that regulate growth, survival, invasion and drug sensitivity of primary tumor cells, including both BCSCs and non-BCSCs. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 102nd Annual Meeting of the American Association for Cancer Research; 2011 Apr 2-6; Orlando, FL. Philadelphia (PA): AACR; Cancer Res 2011;71(8 Suppl):Abstract nr 1431. doi:10.1158/1538-7445.AM2011-1431