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