Comparison of single-cell RNA-seq methods to enable transcriptome profiling of neutrophils in clinical samples.

Monitoring neutrophil gene expression is a powerful tool for understanding disease mechanisms, developing diagnostics, enhancing therapies, and optimizing clinical trials. Neutrophils are sensitive to the processing, storage, and transportation steps that are involved in clinical sample analysis. This study evaluates the capabilities of technologies from 10× Genomics, PARSE Biosciences, and HIVE (Honeycomb Biotechnologies) to generate single-cell RNA sequencing (scRNA-seq) data from human blood-derived neutrophils. Our comparative analysis shows that all methods produced high-quality data, importantly capturing the transcriptomes of neutrophils. Here, we establish a reliable scRNA-seq workflow for neutrophils in clinical trials: we offer guidelines on sample collection to preserve RNA quality and demonstrate how each method performs in capturing sensitive cell populations in clinical practice.