Fully Automated One-Step Production of Functional 3D Tumor Spheroids for High-Content Screening

Three-dimensional culture systems in cancer research: Focus on tumor spheroid model

“…In direct photo patterning, UV exposure produces reactive oxygen species (ROS), which in turn makes the protein-repellent part of a molecule that has been grafted on the substrate to detach, allowing ECM protein to further bind onto the substrate (Théry, 2010). One advantage of direct photo patterning is that it does not require etching, like micro-contact printing (Monjaret et al , 2015). Some of the drawbacks of micropatterning include unequal protein transfer onto the substrate during stamping, requirement of dedicated chemistry to engineer photosensitive materials, and use of bio-incompatible photosensitizers.…”

A High‐Throughput Automated Confocal Microscopy Platform for Quantitative Phenotyping of Nanoparticle Uptake and Transport in Spheroids

“…This strategy is suitable for in situ immunostaining by promoting retention of the 3D structures in the wells, and has been exploited to measure morphological drug responses in 3D tumor spheroid models, in an image‐based high‐throughput manner . We decided to use HT‐29 human colorectal cancer cells as a model previously characterized to be amenable for formation of mature spheroids after a few days of growth, and apply a 4 day culture period to best combine assembly of spheroids with a protocol for RNA interference (RNAi). In the absence of MTG the HT‐29 cells formed complete monolayers, however in the presence of MTG after the same incubation period, we observed the formation of numerous compact spheroids of small and medium sizes per well.…”

“…[8][9][10][11]32] A larger number of spheroids can be assayed in micropatterned-well plates or microfluidic chips. [33][34][35][36] These devices allow in situ immunofluorescence and scalable imaging; however often require specialized fabrication and can be highcost. It therefore remains necessary to refine approaches that will permit the interrogation of phenotypes at the subcellular level but in very high numbers of spheroids in the most technically straightforward manner.…”

Cell spheroids: the new frontiers in in vitro models for cancer drug validation

“…Because drug high-content screening is of great significance in the development of anticancer agents, an automated one-step production method was used for the formation of nine uniform spheroids per well on micropatterned 96-well plates, and validation of fluorouracil, irinotecan and staurosporine in HT-29 spheroids was performed. Micropatterned spheroid imaging shows characteristic cell heterogeneity with distinct microregions, whereas central necrosis appears at a consistent spheroid size, suggesting standardized growth [63].…”