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Single-Cell mRNA Sequencing of the Mouse Brain Vasculature

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Lymphangiogenesis

Part of the book series: Methods in Molecular Biology ((MIMB,volume 1846))

Abstract

In this chapter, we describe a method for analyzing the vasculature of the mouse brain using single-cell transcriptomics. More specifically, we focus on the use of fluorescence-activated cell sorting (FACS) for selection of the cells of interest and sorting of these cells in a 384-well format, allowing for enrichment for the cells being studied. Furthermore, we outline the Smart-Seq2 single-cell library construction method for transforming single-cell mRNA into Illumina sequencing compatible libraries. As single-cell sequencing is still a costly technology, we take special care to describe strategies to include many quality control steps. Finally, we touch upon techniques to convert the sequencing data into a meaningful biological readout. The methods reported in this chapter can be expanded toward other tissues and will prove useful also for the study of different cell types beyond adult brain vasculature.

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Author information

Authors and Affiliations

  1. Karolinska Institutet/AstraZeneca Integrated Cardio Metabolic Centre (KI/AZ ICMC), Huddinge, Sweden

    Michael Vanlandewijck & Christer Betsholtz

  2. Department of Immunology, Genetics and Pathology, Rudbeck Laboratory, Uppsala University, Uppsala, Sweden

    Michael Vanlandewijck & Christer Betsholtz

Authors
  1. Michael Vanlandewijck
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  2. Christer Betsholtz
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Corresponding author

Correspondence to Michael Vanlandewijck .

Editor information

Editors and Affiliations

  1. Center for Vascular & Developmental Biology, Feinberg Cardiovascular Research Institute, Northwestern University, Chicago, IL, USA

    Guillermo Oliver

  2. Department of Medicine and Cardiovascular Institute, University of Pennsylvania, Philadelphia, PA, USA

    Mark L. Kahn

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Vanlandewijck, M., Betsholtz, C. (2018). Single-Cell mRNA Sequencing of the Mouse Brain Vasculature. In: Oliver, G., Kahn, M. (eds) Lymphangiogenesis. Methods in Molecular Biology, vol 1846. Humana Press, New York, NY. https://doi.org/10.1007/978-1-4939-8712-2_21

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  • DOI: https://doi.org/10.1007/978-1-4939-8712-2_21

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  • Publisher Name: Humana Press, New York, NY

  • Print ISBN: 978-1-4939-8711-5

  • Online ISBN: 978-1-4939-8712-2

  • eBook Packages: Springer Protocols

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