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