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

2020; eLife Sciences Publications Ltd; Volume: 9; Linguagem: Inglês

10.7554/elife.52904

2050-084X

Henrik Renner, Martha Grabos, Katharina Becker, Theresa 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,

Single-cell and spatial transcriptomics

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.