| Autor: |
Henrik Renner, Martha Grabos, Katharina J Becker, Theresa E Kagermeier, Jie Wu, Mandy Otto, Stefan Peischard, Dagmar Zeuschner, Yaroslav TsyTsyura, Paul Disse, Jürgen Klingauf, Sebastian A Leidel, Guiscard Seebohm, Hans R Schöler, Jan M Bruder |
| Jazyk: |
angličtina |
| Rok vydání: |
2020 |
| Předmět: |
|
| Zdroj: |
eLife, Vol 9 (2020) |
| Druh dokumentu: |
article |
| ISSN: |
2050-084X |
| DOI: |
10.7554/eLife.52904 |
| Popis: |
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. |
| Databáze: |
Directory of Open Access Journals |
| Externí odkaz: |
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