384 hanging drop arrays give excellent Z ‐factors and allow versatile formation of co‐culture spheroids

Abstract

We previously reported the development of a simple, user‐friendly, and versatile 384 hanging drop array plate for 3D spheroid culture and the importance of utilizing 3D cellular models in anti‐cancer drug sensitivity testing. The 384 hanging drop array plate allows for high‐throughput capabilities and offers significant improvements over existing 3D spheroid culture methods. To allow for practical 3D cell‐based high‐throughput screening and enable broader use of the plate, we characterize the robustness of the 384 hanging drop array plate in terms of assay performance and demonstrate the versatility of the plate. We find that the 384 hanging drop array plate performance is robust in fluorescence‐ and colorimetric‐based assays through Z ‐factor calculations. Finally, we demonstrate different plate capabilities and applications, including: spheroid transfer and retrieval for Janus spheroid formation, sequential addition of cells for concentric layer patterning of different cell types, and culture of a wide variety of cell types. Biotechnol. Bioeng. 2012; 109:1293–1304. © 2011 Wiley Periodicals, Inc. This paper characterizes the robustness of the high‐throughput 384 hanging drop array spheroid formation and culture plate in terms of assay performance. The versatility of the platform was further demonstrated through 3D patterning of multiple cell types into concentric layers and as Janus spheroids. The system is envisioned to deliver valuable insights into 3D cellular behavior as well as more accurate readouts from 3D cell‐based high‐throughput screening and testing.