High-Content Monitoring of Drug Effects in a 3D Spheroid Model

Abstract

A recent decline in the discovery of novel medications challenges the widespread use of 2D monolayer cell assays in the drug discovery process. As a result, the need for more appropriate cellular models of human physiology and disease has renewed the interest in spheroid 3D culture as a pertinent model for drug screening. However, despite technological progress that has significantly simplified spheroid production and analysis, the seeming complexity of the 3D approach has delayed its adoption in many laboratories. The present report demonstrates that the use of a spheroid model may be straightforward and can provide information that is not directly available with a standard 2D approach. We describe a cost-efficient method that allows for the production of an array of uniform spheroids, their staining with vital dyes, real-time monitoring of drug effects, and an ATP-endpoint assay, all in the same 96-well U-bottom plate. To demonstrate the method performance, we analyzed the effect of the preclinical anticancer drug MLN4924 on spheroids formed by VCaP and LNCaP prostate cancer cells. The drug has different outcomes in these cell lines, varying from cell cycle arrest and protective dormancy to senescence and apoptosis. We demonstrate that by using high-content analysis of spheroid arrays, the effect of the drug can be described as a series of EC50 values that clearly dissect the cytostatic and cytotoxic drug actions. The method was further evaluated using four standard cancer chemotherapeutics with different mechanisms of action, and the effect of each drug is described as a unique multi-EC50 diagram. Once fully validated in a wider range of conditions, this method could be particularly valuable for phenotype-based drug discovery.

A Spheroid Killing Assay by CAR T Cells

“…This method is so far only limited to a certain type of adherent cell line (in this publication HCT 116); further development is required to generalize this protocol to every cancerous cell line. Although a wide range of methods is already available for a greater number of target cells 12,13,14,15 , most of them rely on the use of expensive and/or complex procedures. Our method, although limited to a few types of target cells so far, is interesting as it requires very limited inputs from the experimenter and permits a fast and easy screen of various drugs and/or immunotherapy treatments (here CAR CD19 cells).…”

Nanoparticle encapsulation of HDACi with HA targeting in endometrial cancer models

“…Adopting a spheroid model can provide information such as spheroid assembly and morphology, that is not directly available with a standard 2D approach [71]. Spheroid image ( Fig.…”

Review on biofabrication and applications of heterogeneous tumor models

“…Spheroids in gels or scaffolds have also been used for drug testing (Yip & Cho, ). The effect of the preclinical drug MLN4924 on cell cycle arrest was analyzed by tumor spheroids models in Mittler et al's () study. Mi, Liu, Du, Yi, and Sun () studied the cowork of macrophages and paclitaxel on breast cancer cells through a microfluidic system.…”