Microscale screening systems for 3D cellular microenvironments: platforms, advances, and challenges

Abstract

The increasing interest in studying cells using more in vivo-like three-dimensional (3D) microenvironments has created a need for advanced 3D screening platforms with enhanced functionalities and increased throughput. 3D screening platforms that better mimic in vivo microenvironments with enhanced throughput would provide more in-depth understanding of the complexity and heterogeneity of microenvironments. The platforms would also better predict the toxicity and efficacy of potential drugs in physiologically relevant conditions. Traditional 3D culture models (e.g. spinner flasks, gyratory rotation devices, non-adhesive surfaces, polymers) were developed to create 3D multicellular structures. However, these traditional systems require large volumes of reagents and cells, and are not compatible with high throughput screening (HTS) systems. Microscale technology offers the miniaturization of 3D cultures and allows efficient screening of various conditions. This review will discuss the development, most influential works, and current advantages and challenges of microscale culture systems for screening cells in 3D microenvironments.

Advances in multicellular spheroids formation

“…Furthermore, integration of analytical tools with microfluidics ( figure 6), such as Raman, fluorescence spectrometry and UV-Vis spectrometry, enables a rapid, in situ and dynamical analysis during MCS formation. To screen optimized culture conditions for forming MCSs from rare cells or primary cells the microfluidics, consumption of chemical or biochemical reagents is reduced since microfluidics often requires a very small amount of liquid [108]. Furthermore, oxygen supply to MCSs can be significantly improved through microfluidic chips.…”

Beta-hairpin hydrogels as scaffolds for high-throughput drug discovery in three-dimensional cell culture

“…However, most cell-based HTS is done using well-established two-dimensional (2D) cultures which often fail to predict in vivo efficacy of candidate compounds, thereby contributing to the high failure rate and cost of drug development. This is particularly true for cell line models of cancer [1]. While 2D cultures are more convenient and can be automated easily, 3D systems better mimic microenvironments including concentration gradients of nutrients and oxygen and mechanical cues, both of which are particularly important in tumor development [1–3].…”

“…This is particularly true for cell line models of cancer [1]. While 2D cultures are more convenient and can be automated easily, 3D systems better mimic microenvironments including concentration gradients of nutrients and oxygen and mechanical cues, both of which are particularly important in tumor development [1–3]. Due to these differences, cells grown in 3D respond differently to stimuli and drug treatments, and increasing use of 3D cultures as a platform for drug discovery is expected to result in more candidate compounds with high in vivo efficacy and potential for future preclinical studies and clinical drug development.…”

Reconfigurable microfluidic device with discretized sidewall

“…As described in recent reviews, [1][2][3][4][5][6][7][8] many advanced cell culture systems, such as single cell culture, coculture, and 3D culture, have been developed, in addition to microfluidic cell culture systems for in vivo-like perfusion culture. To accomplish complex handling of flows, cells, and cell culture surfaces in microfluidic systems, various microsized structures have been used, including meanders, 9 pillars, 10 branching and confluence of microchannels for cell sorting, 11 stimulation, 12 patterning, 13 and traps.…”