Spheroid arrays for high-throughput single-cell analysis of spatial patterns and biomarker expression in 3D

Abstract

We describe and share a device, methodology and image analysis algorithms, which allow up to 66 spheroids to be arranged into a gel-based array directly from a culture plate for downstream processing and analysis. Compared to processing individual samples, the technique uses 11-fold less reagents, saves time and enables automated imaging. To illustrate the power of the technology, we showcase applications of the methodology for investigating 3D spheroid morphology and marker expression and for in vitro safety and efficacy screens. First, spheroid arrays of 11 cell-lines were rapidly assessed for differences in spheroid morphology. Second, highly-positive (SOX-2), moderately-positive (Ki-67) and weakly-positive (βIII-tubulin) protein targets were detected and quantified. Third, the arrays enabled screening of ten media compositions for inducing differentiation in human neurospheres. Last, the application of spheroid microarrays for spheroid-based drug screens was demonstrated by quantifying the dose-dependent drop in proliferation and increase in differentiation in etoposide-treated neurospheres.

3D tumor spheroids as in vitro models to mimic in vivo human solid tumors resistance to therapeutic drugs

“…Fang & Eglen, 2017). Nonetheless, over the years, the number of research papers that describe new methodologies and technologies for the analysis of the spheroids has been increasing exponentially (Ivanov & Grabowska, 2017;Rodrigues et al, 2018). Nonetheless, over the years, the number of research papers that describe new methodologies and technologies for the analysis of the spheroids has been increasing exponentially (Ivanov & Grabowska, 2017;Rodrigues et al, 2018).…”

“…In fact, despite some studies point out that the results obtained from the therapeutics screening in spheroids be similar to those of in vivo models (Gunness et al, 2013), still remains to be proved the correlation between therapeutics efficacy in spheroids and the results obtained during clinical trials. Nonetheless, over the years, the number of research papers that describe new methodologies and technologies for the analysis of the spheroids has been increasing exponentially (Ivanov & Grabowska, 2017;Rodrigues et al, 2018).…”

The Healthy and Diseased Microenvironments Regulate Oligodendrocyte Properties

“…However, upscaling to high throughput seems possible with the introduction of multiwell-based platforms, including those for 3D culture. 148 Because superresolution microscopy has been evolving for years, the ability of screening complex cultures at high resolution will make imaging of different OL-microenvironmental interactions possible. Because myelin is necessary for the efficient and rapid transmission of action potentials, properties of neuronal activity are another way to analyze (re)generation of myelin.…”

3D tumour spheroids for the prediction of the effects of radiation and hyperthermia treatments

“…But it remains to be shown how this effect translates into spheroid growth response where cells remain in the spheroid microenvironment after treatment rather than being disaggregated and plated at (non-physiological) clonal densities. Although spheroid cultures had already been introduced in the 1970's 19 , recent advances in high-throughput analysis using micro-well spheroid arrays and automated imaging now enable larger scale studies [20][21][22][23] .…”