A fully automated high-throughput workflow for 3D-based chemical screening in human midbrain organoids
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- Henrik Renner
- Department for Cell and Developmental Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany
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- Martha Grabos
- Department for Cell and Developmental Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany
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- Katharina J Becker
- Department for Cell and Developmental Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany
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- Theresa E Kagermeier
- Department for Cell and Developmental Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany
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- Jie Wu
- Max Planck Research Group for RNA Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany
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- Mandy Otto
- Department for Cell and Developmental Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany
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- Stefan Peischard
- Department of Cardiovascular Medicine, Institute for Genetics of Heart Diseases, University Hospital Münster, Münster, Germany
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- Dagmar Zeuschner
- Electron Microscopy Unit, Max Planck Institute for molecular Biomedicine, Münster, Germany
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- Yaroslav TsyTsyura
- Cellular Biophysics Group, Institute for Medical Physics and Biophysics, Westfälische Wilhelms-Universität Münster, Münster, Germany
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- Paul Disse
- Department of Cardiovascular Medicine, Institute for Genetics of Heart Diseases, University Hospital Münster, Münster, Germany
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- Jürgen Klingauf
- Cellular Biophysics Group, Institute for Medical Physics and Biophysics, Westfälische Wilhelms-Universität Münster, Münster, Germany
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- Sebastian A Leidel
- Max Planck Research Group for RNA Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany
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- Guiscard Seebohm
- Department of Cardiovascular Medicine, Institute for Genetics of Heart Diseases, University Hospital Münster, Münster, Germany
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- Hans R Schöler
- Department for Cell and Developmental Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany
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- Jan M Bruder
- Department for Cell and Developmental Biology, Max Planck Institute for molecular Biomedicine, Münster, Germany
説明
<jats:p>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.</jats:p>
収録刊行物
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- eLife
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eLife 9 e52904-, 2020-11-03
eLife Sciences Publications, Ltd