In JoVE (1)
Other Publications (1)
Articles by Darcy Fyffe in JoVE
Robotic Production of Cancer Cell Spheroids with an Aqueous Two-phase System for Drug Testing Stephanie Lemmo Ham1, Ehsan Atefi1, Darcy Fyffe1, Hossein Tavana1 1Department of Biomedical Engineering, The University of Akron A protocol for robotic printing of cancer cell spheroids in a high throughput 96-well plate format using an aqueous two-phase system is presented.
Other articles by Darcy Fyffe on PubMed
High Throughput, Polymeric Aqueous Two-Phase Printing of Tumor Spheroids Advanced Functional Materials. Nov, 2014 | Pubmed ID: 25411577 This paper presents a new 3D culture microtechnology for high throughput production of tumor spheroids and validates its utility for screening anti-cancer drugs. We use two immiscible polymeric aqueous solutions and microprint a submicroliter drop of the "patterning" phase containing cells into a bath of the "immersion" phase. Selecting proper formulations of biphasic systems using a panel of biocompatible polymers results in the formation of a round drop that confines cells to facilitate spontaneous formation of a spheroid without any external stimuli. Adapting this approach to robotic tools enables straightforward generation and maintenance of spheroids of well-defined size in standard microwell plates and biochemical analysis of spheroids in situ, which is not possible with existing techniques for spheroid culture. To enable high throughput screening, we establish a phase diagram to identify minimum cell densities within specific volumes of the patterning drop to result in a single spheroid. Spheroids show normal growth over long-term incubation and dose-dependent decrease in cellular viability when treated with drug compounds, but present significant resistance compared to monolayer cultures. The unprecedented ease of implementing this microtechnology and its robust performance will benefit high throughput studies of drug screening against cancer cells with physiologically-relevant 3D tumor models.