Generation and downstream analysis of single-cell and single-nuclei transcriptomes in brain organoids

Item Type:	Article
Title:	Generation and downstream analysis of single-cell and single-nuclei transcriptomes in brain organoids
Creators Name:	Wandres, M. and Aigner, D. and Kastelic, N. and Boltengagen, A. and Rybak-Wolf, A. and Rajewsky, N.
Abstract:	Over the past decade, single-cell transcriptomics has significantly evolved and become a standard laboratory method for simultaneous analysis of gene expression profiles of individual cells, allowing the capture of cellular diversity. In order to overcome limitations posed by difficult-to-isolate cell types, an alternative approach aiming at recovering single nuclei instead of intact cells can be utilized for sequencing, making transcriptome profiling of individual cells universally applicable. These techniques have become a cornerstone in the study of brain organoids, establishing them as models of the developing human brain. Leveraging the potential of single-cell and single-nucleus transcriptomics in brain organoid research, this protocol presents a step-by-step guide encompassing key procedures such as organoid dissociation, single-cell or nuclei isolation, library preparation and sequencing. By implementing these alternative approaches, researchers can obtain high-quality datasets, enabling the identification of neuronal and non-neuronal cell types, gene expression profiles, and cell lineage trajectories. This facilitates comprehensive investigations into cellular processes and molecular mechanisms shaping brain development.
Keywords:	Brain, Cell Nucleus, Gene Expression Profiling, Organoids, Transcriptome
Source:	Journal of Visualized Experiments
ISSN:	1940-087X
Publisher:	JoVE
Number:	205
Page Range:	e66225
Date:	29 March 2024
Official Publication:	https://doi.org/10.3791/66225
PubMed:	View item in PubMed

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