Abstract
Accumulating evidence indicates that the immune system is regulated not only by immune cells but also by stromal cells in the tissue microenvironment. Characterization of non-hematopoietic cells has not been performed in depth, since markers of the subsets are limited. Recent advances of single-cell technology allow researchers to characterize comprehensively the heterogeneity of stromal cells in an unbiased manner. In this article, we provide step-by-step protocols for cell preparation for single-cell RNA sequencing to characterize the heterogeneity of stroma in human lymph nodes. For complete details on the use and execution of this protocol, please refer to Takeda et al. (2019).