Abstract
Single-cell transcriptomics is a powerful tool to study previously unrealized cellular heterogeneity at the resolution of individual cells. Most of the previous knowledge in cell biology is based on data generated by bulk analysis methods, which provide averaged readouts that usually mask cellular heterogeneity. This approach is challenging when the biological effect of interest is limited to a subpopulation within a cell type. This may particularly apply immune cell populations as these cells are highly mobile and swiftly respond to changes in cytokines or chemokines. For example, in cancer certain subset of myeloid immune cells may acquire immunosuppressive features to suppress antitumor immune responses, and thus described as myeloid-derived suppressor cells (MDSCs). Advances in single-cell RNA sequencing (scRNAseq) allowed scientists to overcome this limitation and enable in-depth interrogation of these subsets of immune cells including MDSCs. Here, we provide a detailed protocol for using scRNAseq to explore MDSCs in the context of splenic myeloid cells from breast tumor-bearing mice in comparison to wildtype controls to define the unique molecular features of immunosuppressive myeloid cells.
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References
Wagner A, Regev A, Yosef N et al (2016) Revealing the vectors of cellular identity with single-cell genomics. Nat Biotechnol 34:1145–1160
Nguyen QH, Pervolarakis N et al (2018) Experimental considerations for single-cell RNA sequencing approaches. Front Cell Dev Biol 6:1–7
Alshetaiwi H, Pervolarakis N et al (2020) Defining the emergence of myeloid-derived suppressor cells in breast cancer using single-cell transcriptomics. Sci Immunol 5:eaay6017
Bronte V, Brandau S et al (2016) Recommendations for myeloid-derived suppressor cell nomenclature and characterization standards. Nat Commun 7:1–10
Solito S, Pinton L et al (2019) Methods to measure MDSC immune suppressive activity in vitro and in vivo. Curr Protoc Immunol 124:e61
Acknowledgments
This study was supported by funding from NIH/NCI (1R01CA234496 and 4R00CA181490 to K.K.); the American Cancer Society (132551-RSG-18-194-01-DDC to K.K.), and the University of Hail, Hail, Saudi Arabia for the Ph.D. Fellowship (to H.A.).
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Alshetaiwi, H., Pervolarakis, N., Nguyen, Q.H., Kessenbrock, K. (2021). Single-Cell Transcriptome Analysis Workflow for Splenic Myeloid-Derived Suppressor Cells from Murine Breast Cancer Models. In: Brandau, S., Dorhoi, A. (eds) Myeloid-Derived Suppressor Cells. Methods in Molecular Biology, vol 2236. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-1060-2_14
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DOI: https://doi.org/10.1007/978-1-0716-1060-2_14
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Publisher Name: Humana, New York, NY
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Online ISBN: 978-1-0716-1060-2
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