![]() ![]() Harnessing the benefits of the standard 96-well plate format, 3D spheroidal cultures with compatible designs were developed to address several applications. The practical utilization of a 96-well plate for biological inquiry enables the execution of parallel and large-scale independent experiments. The conceptualized WPF approach can be adapted for a range of applications in both basic and applied biological/engineering research. As a proof-of-concept, we also demonstrate matrix-assisted tumor culture techniques in advancing the broader use of 3D culture systems. To demonstrate the applicability and practicality of the conceived approach, we utilized human colorectal carcinoma cells (HCT116) to first show the generation of large scaffold-free 3D tumor spheroids over 1.5 mm in diameter in single-well plates. The droplets of the medium generated by the well-plate flip (WPF) method can be easily modified to address various mechanisms and processes in cell biology, including cancer. Hanging drop preparation can be easily employed by flipping a universal 96-well plate. In this study, we introduce a straightforward and user-friendly approach based on standard 96-well plates with basic amenities for growing 3D tumors in a scaffold-free/scaffold-based format. This, in turn, limits their applicability and standardization. Despite the widespread interest, need, and present-day effort, most of the 3D tumor culturing methodologies have not gone beyond the inventors’ laboratories. Three-dimensional (3D) tumor culture techniques are gaining popularity as in vitro models of tumoral tissue analogues. Woese Institute for Genomic Biology, University of Illinois at Urbana-Champaign, Urbana, IL, United States 3Department of Computer Science, University of Illinois at Urbana‐Champaign, Urbana, IL, United States.2Cancer Center at Illinois, University of Illinois at Urbana-Champaign, Urbana, IL, United States. ![]() 1Department of Bioengineering, University of Illinois at Urbana‐Champaign, Urbana, IL, United States.Yoon Jeong 1,2 Ashley Tin 3 Joseph Irudayaraj 1,2,4* ![]()
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