RESEARCH ARTICLE


Live Multicellular Tumor Spheroid Models For High-Content Imaging and Screening In Cancer Drug Discovery



Brian G. Reid1, Taleen Jerjian1, Purvi Patel2, Qiong Zhou1, Byong Hoon Yoo1, Peter Kabos3, Carol A. Sartorius2, Daniel V. LaBarbera1, *
1 Department of Pharmaceutical Sciences, Skaggs School of Pharmacy and Pharmaceutical Sciences, The University of Colorado, Anschutz Medical Campus, Aurora CO, USA
2 Pathology, School of Medicine, The University of Colorado, Anschutz Medical Campus, Aurora CO, USA
3 Medical Oncology, School of Medicine, The University of Colorado, Anschutz Medical Campus, Aurora CO, USA


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© Reid et al.; Licensee Bentham Open.

open-access license: This is an open access article licensed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted, non-commercial use, distribution and reproduction in any medium, provided the work is properly cited.

* Address correspondence to this author at the Department of Pharmaceutical Sciences, The Skaggs School of Pharmacy and Pharmaceutical Sciences, The University of Colorado, Anschutz Medical Campus, 12850 East Montview Blvd, Room V20-2101, Aurora, CO 80045, USA; Tel: (303)724-4116; Fax: (303)724-7266; E-mail: daniel.labarbera@ucdenver.edu,


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.

Keywords: Multicellular Tumor Spheroid (MCTS), High-Throughput Screening (HTS), High-Content Screening (HCS), Cancer Stem Cells (CSC), Cytokeratin 5, “Systems Biology”, Luminal Breast Cancer, Cancer Drug Discovery.