Gentle cell sorting of immune cells for downstream single-cell RNA-seq

Abstract Cell sorters have played an instrumental role in the progression of immunological discoveries. The ability to isolate specific immune cell types from a heterogenous population while maintaining high viability is critical to the discovery and generation of disease therapies. To isolate fragile and rare immune cell types, a gentle low pressure cell sorter is necessary. In this study, regulatory T cells (Tregs) were sorted from peripheral blood mononuclear cells using the WOLF® Cell Sorter developed by NanoCellect® and a traditional jet-in-air cell sorter. The isolated cells were then analyzed for single-cell RNA-seq using the 10x Genomics workflow. We also analyzed how different sorting methods affect the expression of stress markers induced during sorting. This study showed that despite the small percentage of Tregs in peripheral blood mononuclear cells, the WOLF Cell Sorter is able to obtain high purity, high viability, and high quality sequencing results. The WOLF Cell Sorter was also able analyze several subsets within the purified Treg populations in higher definition. Furthermore, fewer stress markers were induced when compared to the traditional jet-in-air cell sorter. This study provides novel insights into a robust and simplified workflow to use when isolating fragile immune cell types.