Development of a miniaturized 3D organoid culture platform for ultra-high-throughput screening.

Journal of Molecular Cell Biology
Yuhong DuHaian Fu

Abstract

The recent advent of robust methods to grow human tissues as 3D organoids allows us to recapitulate the 3D architecture of tumors in an in vitro setting and offers a new orthogonal approach for drug discovery. However, organoid culturing with extracellular matrix to support 3D architecture has been challenging for high-throughput screening (HTS)-based drug discovery due to technical difficulties. Using genetically engineered human colon organoids as a model system, here we report our effort to miniaturize such 3D organoid culture with extracellular matrix support in high-density plates to enable HTS. We first established organoid culturing in a 384-well plate format and validated its application in a cell viability HTS assay by screening a 2036-compound library. We further miniaturized the 3D organoid culturing in a 1536-well ultra-HTS format and demonstrated its robust performance for large-scale primary compound screening. Our miniaturized organoid culturing method may be adapted to other types of organoids. By leveraging the power of 3D organoid culture in a high-density plate format, we provide a physiologically relevant screening platform to model tumors to accelerate organoid-based research and drug discovery.

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Citations

Nov 24, 2020·Cell Death and Differentiation·Therese SeidlitzDaniel E Stange
Apr 14, 2021·NPJ Precision Oncology·G Emerens WensinkJeanine M L Roodhart
Jun 22, 2021·Advanced Drug Delivery Reviews·Zhimin LuoAli Khademhosseini
Jun 19, 2021·Nature Cancer·Yuan-Hung LoCalvin J Kuo

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Methods Mentioned

BETA
xenograft
genetic modification
electrophoresis

Software Mentioned

GraphPad Prism
CambridgeSoft Bioassay
ImageXpress
Cambridge BioAssay
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