title: Live multicellular tumor spheroid models for high-content imaging and screening in cancer drug discovery.
abstract: The multi cellular tumor spheroid (MCTS) model has been used for decades with proven superiority over monolayer cell culture models at recapitulating in vivo tumor growth. Yet its use in high-throughput drug discovery has been limited, particularly with image based screening, due to practical and technical hurdles. Here we report a significant advance in utilizing live MCTS models for high-content image based drug discovery. Using a validated GFP reporter (CK5Pro-GFP) of luminal breast cancer stem cells (CSC), we developed an algorithm to quantify changes in CK5Pro-GFP expression levels for individual Z-stack planes (local) or as maximal projections of the summed Z-stacks (global) of MCTS. From these image sets, we can quantify the cross-sectional area of GFP positive cells, the fluorescence intensity of the GFP positive cells, and the percent of spheroid cross-sectional area that expresses CK5Pro-GFP.We demonstrate that acquiring data in this manner can be done in real time and is statistically robust (Z'=0.85) for use in primary high-content screening cancer drug discovery.
authors.affiliation: Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, The University of Colorado, Anschutz Medical Campus, Aurora CO, USA.
authors.forename: Brian G
authors.initials: BG
authors.lastname: Reid
authors.affiliation: Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, The University of Colorado, Anschutz Medical Campus, Aurora CO, USA.
authors.forename: Taleen
authors.initials: T
authors.lastname: Jerjian
authors.affiliation: Pathology, School of Medicine, The University of Colorado, Anschutz Medical Campus, Aurora CO, USA.
authors.forename: Purvi
authors.initials: P
authors.lastname: Patel
authors.affiliation: Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, The University of Colorado, Anschutz Medical Campus, Aurora CO, USA.
authors.forename: Qiong
authors.initials: Q
authors.lastname: Zhou
authors.affiliation: Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, The University of Colorado, Anschutz Medical Campus, Aurora CO, USA.
authors.forename: Byong Hoon
authors.initials: BH
authors.lastname: Yoo
authors.affiliation: Medical Oncology, School of Medicine, The University of Colorado, Anschutz Medical Campus, Aurora CO, USA.
authors.forename: Peter
authors.initials: P
authors.lastname: Kabos
authors.affiliation: Pathology, School of Medicine, The University of Colorado, Anschutz Medical Campus, Aurora CO, USA.
authors.forename: Carol A
authors.initials: CA
authors.lastname: Sartorius
authors.affiliation: Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, The University of Colorado, Anschutz Medical Campus, Aurora CO, USA.
authors.forename: Daniel V
authors.initials: DV
authors.lastname: Labarbera
keywords: Cancer Drug Discovery.; Cancer Stem Cells (CSC); Cytokeratin 5; High-Content Screening (HCS); High-Throughput Screening (HTS); Luminal Breast Cancer; Multicellular Tumor Spheroid (MCTS); “Systems Biology”