Department of Biological Chemistry, University of California, Irvine, Irvine, CA, USA.
Department of Pathology, University of Hail, Hail, Saudi Arabia.
Methods Mol Biol. 2021;2236:177-187. doi: 10.1007/978-1-0716-1060-2_14.
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.
单细胞转录组学是一种强大的工具，可以在单个细胞的分辨率下研究以前未实现的细胞异质性。大多数以前的细胞生物学知识都是基于批量分析方法生成的数据，这些数据提供了平均读数，通常掩盖了细胞异质性。当感兴趣的生物学效应仅限于细胞类型中的亚群时，这种方法具有挑战性。这可能特别适用于免疫细胞群体，因为这些细胞具有高度的移动性，并能迅速响应细胞因子或趋化因子的变化。例如，在癌症中，某些髓系免疫细胞亚群可能获得免疫抑制特征，以抑制抗肿瘤免疫反应，因此被描述为髓源性抑制细胞（MDSC）。单细胞 RNA 测序（scRNAseq）的进展使科学家能够克服这一限制，并能够深入研究这些免疫细胞亚群，包括 MDSC。在这里，我们提供了一个详细的方案，用于使用 scRNAseq 来探索乳腺癌荷瘤小鼠脾脏髓系细胞中的 MDSC，并与野生型对照进行比较，以定义免疫抑制性髓系细胞的独特分子特征。
