Abstract
Natural killer (NK) cells are innate lymphocytes that control tumors and microbial infections. Human NK cells are transcriptomically and phenotypically heterogeneous. The site where NK cells develop and reside determines their phenotype and effector functions. Our current knowledge about human NK cells is primarily from blood- and bone marrow-derived NK cells. The major limitation in formulating organ-specific clinical therapy is the knowledge gap on how tissue-resident NK cells develop, home, and function. Thus, it is crucial to define the transcriptomic profiles and the transcriptional regulation of tissue-resident NK cells. The major challenges in studying tissue-resident NK cells include their total number and the complexity of the tissue. Additionally, during isolation, keeping them viable and naïve without activation are challenging tasks. Here, we provide methods for isolating and performing transcriptomic analyses of NK cells at the individual cell level. Single-cell RNA sequencing provides a higher resolution of cellular heterogeneity and a better understanding of cell–cell interactions within the microenvironment. Using these methods, we can efficiently identify distinct populations of NK cells in tissues and define their unique transcriptomic profiles.
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Acknowledgments
We dedicate this work to our inspiring colleague Dr. Mathew Riese MD, Ph.D., who passed away young. This work was supported in part by NIH R01 AI102893; NCI R01 CA179363 (S.M.); HRHM Program of MACC Fund (S.M.), Nicholas Family Foundation (S.M.); Gardetto Family (S.M.); MCW-Cancer Center-Large Seed Grant (S.M.); MACC Fund (S.M.); Ann’s Hope Melanoma Foundation (S.M.); and Advancing Healthier Wisconsin (S.M.).
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Hashemi, E., Mei, A., Wang, D., Khalil, M., Malarkannan, S. (2022). Methods for Isolating and Defining Single-Cell Transcriptomes of Tissue-Resident Human NK Cells. In: Shimasaki, N. (eds) Natural Killer (NK) Cells. Methods in Molecular Biology, vol 2463. Humana, New York, NY. https://doi.org/10.1007/978-1-0716-2160-8_8
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DOI: https://doi.org/10.1007/978-1-0716-2160-8_8
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