Robotic production of cancer cell spheroids with an aqueous two-phase system for drug testing.
Cancer cell spheroids present a relevant in vitro model of avascular tumors for anti-cancer drug testing applications. A detailed protocol for producing both mono-culture and co-culture spheroids in a high throughput 96-well plate format is described in this work. This approach utilizes an aqueous two-phase system to confine cells into a drop of the denser aqueous phase immersed within the second aqueous phase. The drop rests on the well surface and keeps cells in close proximity to form a single spheroid. This technology has been adapted to a robotic liquid handler to produce size-controlled spheroids and expedite the process of spheroid production for compound screening applications. Spheroids treated with a clinically-used drug show reduced cell viability with increase in the drug dose. The use of a standard micro-well plate for spheroid generation makes it straightforward to analyze viability of cancer cells of drug-treated spheroids with a micro-plate reader. This technology is straightforward to implement both robotically and with other liquid handling tools such as manual pipettes.
Department of Biomedical Engineering, The University of Akron.
Stephanie Lemmo
SL
Ham
Department of Biomedical Engineering, The University of Akron.
Ehsan
E
Atefi
Department of Biomedical Engineering, The University of Akron.
Darcy
D
Fyffe
Department of Biomedical Engineering, The University of Akron; tavana@uakron.edu.
Hossein
H
Tavana
D045744:Cell Line, Tumor; D018920:Coculture Techniques; D004354:Drug Screening Assays, Antitumor / Q000295:instrumentation / Q000379:methods*; D057166:High-Throughput Screening Assays / Q000295:instrumentation / Q000379:methods; D006801:Humans; D009369:Neoplasms / Q000188:drug therapy* / Q000473:pathology*; D012371:Robotics / Q000295:instrumentation / Q000379:methods; D018874:Spheroids, Cellular / Q000187:drug effects* / Q000473:pathology*