Development of Hydrogel-Based 3D Cancer Stem Cell & Tumorsphere Models for HTS

Cancer stem cells (CSCs), commonly referred to as tumor initiating cells (TICs), are thought to be the origin of replicating malignant cells that remain after the primary tumor is removed or eradicated. While the contribution of TICs to cancers of hematopoietic origin is well established, there is a growing body of evidence that supports the presence of TICs within solid tumors. This heterogeneity contributes to the intrinsic resistance of solid tumors to chemotherapeutics and eventually leads to therapeutic failure and patient death. Historically, screening for oncology directed compounds has been performed in 2-dimensional monolayer cultures which fail to replicate the complex architecture and microenvironment of tumors in vivo.

To address this need using prototype manufactured 384-well round bottom clear black plates coated with hydrogel (commonly referred to as ultra-low attachment or ULA plates) we have developed a method to generate single cell type spheroids to accommodate their use in high throughput screening. These cultures were systematically compared to spheres grown in conventional plates and comparative imaging analyses were carried out to assess sphere formation. In addition, using the same plates, we have developed multicellular tumor spheroids (MCTS) composed of various cell types cultured in the same well. Differences in the cellular compositions of these spheroids in response to drugs can be quantified using high content imagers such as the WiScan from Idea-Bio and the High Throughput Flow Cytometer from IntelliCyt. These heterotypic, differentially labeled spheroids are a valuable asset in the generation of cell-based HTS assays capable of identifying molecules that selectively kill TICs and the cells comprising their microenvironment.

Dec 4 2013

48 mins