Department for Cell and Developmental Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany
Martha Grabos
Department for Cell and Developmental Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany
Katharina J Becker
Department for Cell and Developmental Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany; Westfälische Wilhelms-Universität Münster, Münster, Germany
Theresa E Kagermeier
Department for Cell and Developmental Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany; Westfälische Wilhelms-Universität Münster, Münster, Germany
Jie Wu
Max Planck Research Group for RNA Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany; Research Group for RNA Biochemistry, Department of Chemistry and Biochemistry, University of Bern, Bern, Switzerland
Mandy Otto
Department for Cell and Developmental Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany; Westfälische Wilhelms-Universität Münster, Münster, Germany
Max Planck Research Group for RNA Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany; Research Group for RNA Biochemistry, Department of Chemistry and Biochemistry, University of Bern, Bern, Switzerland
Guiscard Seebohm
Department of Cardiovascular Medicine, Institute for Genetics of Heart Diseases, University Hospital Münster, Münster, Germany
Hans R Schöler
Department for Cell and Developmental Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany; Westfälische Wilhelms-Universität Münster, Münster, Germany
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.
