Renner, Henrik; Grabos, Martha; Becker, Katharina J; Kagermeier, Theresa E; Wu, Jie; Otto, Mandy; Peischard, Stefan; Zeuschner, Dagmar; TsyTsyura, Yaroslav; Disse, Paul; Klingauf, Jürgen; Leidel, Sebastian A; Seebohm, Guiscard; Schöler, Hans R; Bruder, Jan M (2020). A fully automated high-throughput workflow for 3D-based chemical screening in human midbrain organoids. eLife, 9 eLife Sciences Publications 10.7554/eLife.52904
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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.
Item Type: |
Journal Article (Original Article) |
|---|---|
Division/Institute: |
08 Faculty of Science > Department of Chemistry, Biochemistry and Pharmaceutical Sciences (DCBP) |
Graduate School: |
Graduate School for Cellular and Biomedical Sciences (GCB) |
UniBE Contributor: |
Wu, Jie, Leidel, Sebastian Andreas |
Subjects: |
500 Science > 570 Life sciences; biology 500 Science > 540 Chemistry |
ISSN: |
2050-084X |
Publisher: |
eLife Sciences Publications |
Language: |
English |
Submitter: |
Christina Schüpbach |
Date Deposited: |
16 Nov 2020 15:16 |
Last Modified: |
08 Jan 2023 22:04 |
Publisher DOI: |
10.7554/eLife.52904 |
BORIS DOI: |
10.7892/boris.148118 |
URI: |
https://boris.unibe.ch/id/eprint/148118 |
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