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

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

  • Renner, Henrik ORCID Department for Cell and Developmental Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany
  • Grabos, Martha Department for Cell and Developmental Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany
  • Becker, Katharina J Westfälische Wilhelms-Universität Münster, Münster, Germany
  • Kagermeier, Theresa E Westfälische Wilhelms-Universität Münster, Münster, Germany
  • Wu, Jie Research Group for RNA Biochemistry, Department of Chemistry and Biochemistry, University of Bern, Bern, Switzerland
  • Otto, Mandy Westfälische Wilhelms-Universität Münster, Münster, Germany
  • Peischard, Stefan ORCID Department of Cardiovascular Medicine, Institute for Genetics of Heart Diseases, University Hospital Münster, Münster, Germany
  • Zeuschner, Dagmar Electron Microscopy Unit, Max Planck Institute for molecular Biomedicine, Münster, Germany
  • TsyTsyura, Yaroslav Cellular Biophysics Group, Institute for Medical Physics and Biophysics, Westfälische Wilhelms-Universität Münster, Münster, Germany
  • Disse, Paul Department of Cardiovascular Medicine, Institute for Genetics of Heart Diseases, University Hospital Münster, Münster, Germany
  • Klingauf, Jürgen Cellular Biophysics Group, Institute for Medical Physics and Biophysics, Westfälische Wilhelms-Universität Münster, Münster, Germany
  • Leidel, Sebastian A ORCID Research Group for RNA Biochemistry, Department of Chemistry and Biochemistry, University of Bern, Bern, Switzerland
  • Seebohm, Guiscard Department of Cardiovascular Medicine, Institute for Genetics of Heart Diseases, University Hospital Münster, Münster, Germany
  • Schöler, Hans R Westfälische Wilhelms-Universität Münster, Münster, Germany
  • Bruder, Jan M ORCID Department for Cell and Developmental Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany
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  • 2020-11-3
Published in:
  • eLife. - eLife Sciences Publications, Ltd. - 2020, vol. 9
General Biochemistry, Genetics and Molecular Biology
General Immunology and Microbiology
General Neuroscience
General Medicine
English 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.
Language
  • English
Open access status
gold
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Persistent URL
https://folia.unifr.ch/global/documents/132032
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