Evaluating the WOLF® Cell Sorter impact on neutrophil viability and activation

Abstract

Abstract Neutrophils have been associated with antimicrobial functions in acute infections and also play critical roles in chronic inflammation. The half-life of circulating neutrophils is estimated at between 13–19 hours and, if activated, they may be cleared even faster. The shear forces and pressure of traditional FACS can be harsh on such sensitive cells and as a result, the use of magnetic beads and centrifugation gradients to isolate neutrophils is sometimes preferred. However, these methods lack the ability to select for viability or multiple markers needed for specific subset gating. NanoCellect’s WOLF Cell Sorter allows for multi-parameter detection and sorting like traditional FACS. However, the WOLF’s microfluidic sorting mechanism uses far lower pressure (<2 psi vs 10–70 psi) that results in gentler sorting and a higher quality of cells post-sort. Here, the ability of this novel instrument to sort neutrophils with minimal cell death or activation was investigated. Live CD16+ neutrophils were sorted from whole blood and evaluated for purity, viability, and expression levels of CD11b, and CD62L. Results demonstrate that the WOLF Cell Sorter can enrich for neutrophils with high purity, viability, and a limited effect on activation, making it an effective method for neutrophil separation. This indicates that this novel microfluidic cell sorting technology can be used to study emerging neutrophil functions, mechanisms of action, and novel cell-based therapeutic approaches.