Figure 1.

Single cell-RNA sequencing can map myeloid cell heterogeneity in the kidney. For dense tissues like the kidney, cell dissociation, typically achieved using both physical disaggregation and enzymatic digestion, is the most important step; it directly affects the molecular profiles of cells and can introduce stress-induced transcriptional artifacts (9,17). This is particularly limiting when looking at kidney disease samples where tissue is subject to inflammatory stress, although this may be overcome by excluding low-quality cells, for example, on the basis of high mitochondrial RNA content (18). Cell isolation and capture is the most unique hurdle to overcome for scRNA-seq. The use of FACS allows for the selection of cells on the basis of surface markers and is, therefore, useful when isolating a specific subset of cells for sequencing, including CD45 for immune cells. Adapted from ref. 10, with permission. R-UUO, reversible unilateral ureteral obstruction; tSNE, t-distributed stochastic neighbor embedding; UUO, unilateral ureteral obstruction.