Isolation of polymorphonuclear neutrophils and monocytes from a single sample of human peripheral blood

Summary Monocytes and neutrophils are widely distributed throughout the body and play essential roles in health and disease. Here, we present a detailed protocol to isolate polymorphonuclear neutrophils and monocytes from a single sample of human peripheral blood. We have optimized several aspects of the procedure, including the density gradient, timing of each cell processing step, and the buffer/media conditions to preserve cell viability for subsequent functional assays. This protocol is reproducible and can be scaled as required for downstream applications. For complete details on the use and execution of this protocol, please refer to Cui et al. (2021).
Graphical abstract
Highlights • A protocol for isolating neutrophils and monocytes from a single human blood sample • Optimizes key processing steps to ensure high purity, yield, and viability of cells • Scalable for a wide range of downstream applications
Monocytes and neutrophils are widely distributed throughout the body and play essential roles in health and disease. Here, we present a detailed protocol to isolate polymorphonuclear neutrophils and monocytes from a single sample of human peripheral blood. We have optimized several aspects of the procedure, including the density gradient, timing of each cell processing step, and the buffer/media conditions to preserve cell viability for subsequent functional assays. This protocol is reproducible and can be scaled as required for downstream applications.