High-throughput 3D spheroid culture and drug testing using a 384 hanging drop array

Abstract

Culture of cells as three-dimensional (3D) aggregates can enhance in vitro tests for basic biological research as well as for therapeutics development. Such 3D culture models, however, are often more complicated, cumbersome, and expensive than two-dimensional (2D) cultures. This paper describes a 384-well format hanging drop culture plate that makes spheroid formation, culture, and subsequent drug testing on the obtained 3D cellular constructs as straightforward to perform and adapt to existing high-throughput screening (HTS) instruments as conventional 2D cultures. Using this platform, we show that drugs with different modes of action produce distinct responses in the physiological 3D cell spheroids compared to conventional 2D cell monolayers. Specifically, the anticancer drug 5-fluorouracil (5-FU) has higher anti-proliferative effects on 2D cultures whereas the hypoxia activated drug commonly referred to as tirapazamine (TPZ) are more effective against 3D cultures. The multiplexed 3D hanging drop culture and testing plate provides an efficient way to obtain biological insights that are often lost in 2D platforms.

Modeling infectious diseases and host-microbe interactions in gastrointestinal organoids

“…Also, in the protected lumen of a 3D structure, conditions may differ from those in 2D layers, for example in regard to O2 levels or the accumulation of mucus or metabolites such as butyrate or urea. These are factors, which could potentially influence the progression of infection, and thus 2D cultures may not provide the level of insight that could be obtained in 3D (Tung et al, 2011).…”

Organ-on-a-Chip Platforms for Drug Delivery and Cell Characterization: A Review

“…(19)(20)(21)(22) The most common 3D cell culturing platforms are cell aggregates, (21) spheroids, (19) and cell sheets. (18,22) Examples of cell aggregates and multicell-type sheets are shown in Figs.…”

Metabolic response of lung cancer cells to radiation in a paper-based 3D cell culture system

“…While 2D cultures are easy to establish experimentally, they fail to mimic many aspects of a tissue because the cells lack the structural features of a tissue: the 3D contacts formed between adjacent cells; 3D contacts between cells and the extracellular matrix; and 3D structural features that limit the mass transport of molecules (e.g., oxygen, glucose, lactate, and carbon dioxide) within the culture environment [38,39]. These factors are crucial to the response of cells to chemo-and radio-therapy [40].…”