Single Cell Methods pp 9-21 | Cite as
Tissue Handling and Dissociation for Single-Cell RNA-Seq
Protocol
First Online:
Abstract
The starting material for all single-cell protocols is a cell suspension. The particular functions and spatial distribution of immune cells generally make them easy to isolate them from the tissues where they dwell. Here we describe tissue dissociation protocols that have been used to obtain human immune cells from lymphoid and nonlymphoid tissues to be then used as input to single-cell methods. We highlight the main factors that can influence the final quality of single-cell data, namely the stress signatures that can bias its interpretation.
Key words
Single-cell RNA sequencing Tissue processing Digestion Single-cell suspension Immune cellsReferences
- 1.van den Brink SC, Sage F, Vértesy Á et al (2017) Single-cell sequencing reveals dissociation-induced gene expression in tissue subpopulations. Nat Methods 14:935–936CrossRefPubMedGoogle Scholar
- 2.Wu YE, Pan L, Zuo Y et al (2017) Detecting activated cell populations using single-cell RNA-seq. Neuron 96:313–329.e6CrossRefPubMedGoogle Scholar
- 3.Adam M, Potter AS, Potter SS (2017) Psychrophilic proteases dramatically reduce single-cell RNA-seq artifacts: a molecular atlas of kidney development. Development 144:3625–3632CrossRefPubMedPubMedCentralGoogle Scholar
- 4.Gunawan M, Jardine L, Haniffa M (2016) Isolation of human skin dendritic cell subsets. Methods Mol Biol 1423:119–128CrossRefPubMedGoogle Scholar
Copyright information
© Springer Science+Business Media, LLC, part of Springer Nature 2019