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Issue 4, 2016
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3D high throughput screening and profiling of embryoid bodies in thermoformed microwell plates

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Abstract

3D organoids using stem cells to study development and disease are now widespread. These models are powerful to mimic in vivo situations but are currently associated with high variability and low throughput. For biomedical research, platforms are thus necessary to increase reproducibility and allow high-throughput screens (HTS). Here, we introduce a microwell platform, integrated in standard culture plates, for functional HTS. Using micro-thermoforming, we form round-bottom microwell arrays from optically clear cyclic olefin polymer films, and assemble them with bottom-less 96-well plates. We show that embryonic stem cells aggregate faster and more reproducibly (centricity, circularity) as compared to a state-of-the-art microwell array. We then run a screen of a chemical library to direct differentiation into primitive endoderm (PrE) and, using on-chip high content imaging (HCI), we identify molecules, including regulators of the cAMP pathway, regulating tissue size, morphology and PrE gene activity. We propose that this platform will benefit to the systematic study of organogenesis in vitro.

Graphical abstract: 3D high throughput screening and profiling of embryoid bodies in thermoformed microwell plates

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Publication details

The article was received on 07 Dec 2015, accepted on 08 Jan 2016 and first published on 08 Jan 2016


Article type: Paper
DOI: 10.1039/C5LC01499A
Citation: Lab Chip, 2016,16, 734-742
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    3D high throughput screening and profiling of embryoid bodies in thermoformed microwell plates

    E. J. Vrij, S. Espinoza, M. Heilig, A. Kolew, M. Schneider, C. A. van Blitterswijk, R. K. Truckenmüller and N. C. Rivron, Lab Chip, 2016, 16, 734
    DOI: 10.1039/C5LC01499A

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