A fully automated high-throughput workflow for 3D-based chemical screening in human midbrain organoids

Renner, Henrik; Grabos, Martha; Becker, Katharina J; Kagermeier, Theresa E; Wu, Jie; Otto, Mandy; Peischard, Stefan; Zeuschner, Dagmar; TsyTsyura, Yaroslav; Disse, Paul; Klingauf, Jürgen; Leidel, Sebastian A; Seebohm, Guiscard; Schöler, Hans R; Bruder, Jan M (2020). A fully automated high-throughput workflow for 3D-based chemical screening in human midbrain organoids. eLife, 9 eLife Sciences Publications 10.7554/eLife.52904

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Three-dimensional (3D) culture systems have fueled hopes to bring about the next generation of more physiologically relevant high-throughput screens (HTS). However, current protocols yield either complex but highly heterogeneous aggregates ('organoids') or 3D structures with less physiological relevance ('spheroids'). Here, we present a scalable, HTS-compatible workflow for the automated generation, maintenance, and optical analysis of human midbrain organoids in standard 96-well-plates. The resulting organoids possess a highly homogeneous morphology, size, global gene expression, cellular composition, and structure. They present significant features of the human midbrain and display spontaneous aggregate-wide synchronized neural activity. By automating the entire workflow from generation to analysis, we enhance the intra- and inter-batch reproducibility as demonstrated via RNA sequencing and quantitative whole mount high-content imaging. This allows assessing drug effects at the single-cell level within a complex 3D cell environment in a fully automated HTS workflow.

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