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Practical Considerations for Complex Tissue Dissociation for Single-Cell Transcriptomics

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Single Cell Transcriptomics

Part of the book series: Methods in Molecular Biology ((MIMB,volume 2584))

Abstract

Single-cell and single-nucleus RNA sequencing have revolutionized biomedical research, allowing analysis of complex tissues, identification of novel cell types, and mapping of development as well as disease states. Successful application of this technology critically relies on the dissociation of solid organs and tissues into high-quality single-cell (or nuclei) suspensions.

In this chapter, we examine several key aspects of the tissue handling workflow that need to be considered when establishing an efficient tissue processing protocol for single-cell RNA sequencing (scRNA-seq). These include tissue collection, transport, and storage, as well as the choice of the dissociation conditions. We emphasize the importance of the tissue quality check and discuss the advantages (and potential limitations) of tissue cryopreservation. We provide practical tips and considerations on each of the steps of the processing workflow, and comment on how to maximize cell viability and integrity, which are critical for obtaining high-quality single-cell transcriptomic data.

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Author information

Authors and Affiliations

  1. Division of Biomedicine, School of Biosciences, Cardiff University, Cardiff, UK

    Stephanie T. Pohl

  2. Division of Cancer Epigenomics, German Cancer Research Center (DKFZ) and Translational Lung Research Center, Heidelberg, Germany

    Maria Llamazares Prada

  3. Anatomy Unit, Department of Pathology and Experimental Therapy, School of Medicine, University of Barcelona (UB), L’Hospitalet de Llobregat, Barcelona, Spain

    Elisa Espinet

  4. School of Biosciences, Cardiff University, Cardiff, Wales, UK

    Renata Jurkowska

Authors
  1. Stephanie T. Pohl
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  2. Maria Llamazares Prada
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  3. Elisa Espinet
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  4. Renata Jurkowska
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Corresponding author

Correspondence to Renata Jurkowska .

Editor information

Editors and Affiliations

  1. Molecular Biotechnology Center, University of Torino, Torino, Italy

    Raffaele A. Calogero

  2. European Molecular Biology Laboratory, Heidelberg, Germany

    Vladimir Benes

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© 2023 The Author(s), under exclusive license to Springer Science+Business Media, LLC, part of Springer Nature

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Pohl, S.T., Prada, M.L., Espinet, E., Jurkowska, R. (2023). Practical Considerations for Complex Tissue Dissociation for Single-Cell Transcriptomics. In: Calogero, R.A., Benes, V. (eds) Single Cell Transcriptomics. Methods in Molecular Biology, vol 2584. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2756-3_19

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  • DOI: https://doi.org/10.1007/978-1-0716-2756-3_19

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  • Publisher Name: Humana, New York, NY

  • Print ISBN: 978-1-0716-2755-6

  • Online ISBN: 978-1-0716-2756-3

  • eBook Packages: Springer Protocols

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