High-throughput drug screening of fine-needle aspiration-derived cancer organoids

Summary

Generation of fine-needle aspiration (FNA)-derived cancer organoids has allowed us to develop a number of downstream applications. In this protocol, we start with organoids cultured in a semi-solid format. We dissociate organoids into single cells and then plate in a 384-well format for high-throughput drug screening. While this method must be fine-tuned for each individual organoid culture, it offers a format well suited for rapidly screening medium-sized drug/compound libraries (500–5,000 molecules) and generating dose-response curves to measure relative efficacy.

For complete details on the use and execution of this protocol, please refer to Lee et al. (2020) and Vilgelm et al. (2020).

Graphical Abstract

Highlights

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  Protocol to dissociate organoids into single cells for high-throughput plating methods

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  Organoids form in a 384-well format when plated in cell-repellent plates

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  Steps for high-throughput drug screening with viability assays

Abstract

Generation of fine-needle aspiration (FNA)-derived cancer organoids has allowed us to develop a number of downstream applications. In this protocol, we start with organoids cultured in a semi-solid format. We dissociate organoids into single cells and then plate in a 384-well format for high-throughput drug screening. While this method must be fine-tuned for each individual organoid culture, it offers a format well suited for rapidly screening medium-sized drug/compound libraries (500–5,000 molecules) and generating dose-response curves to measure relative efficacy.