Simplified Neutrophil Isolation Protocol

Abstract

Background: Neutrophils are the most abundant leukocytes in humans and among the first cells to arrive on the site of inflammatory immune response. Due to their key role in inflammation, neutrophil functions such as locomotion, cytokine production, phagocytosis, respiratory burst and release of extracellular traps are extensively studied. Objective: Develop a simple, fast and cheap method to isolate neutrophils from human blood. Methods: A blood sample of 15 ml was obtained from healthy subjects. Subsequently, neutrophils where obtained using the modified density gradient separation method. Finally cell viability and morphology was analysed. Results: An average of 1.5 × 107 cells/ml was obtained with a > 98% neutrophil purity and a viability greater than 95%. Conclusions: Our protocol demonstrates that neutrophils can be isolated without investing a lot of money per procedure, which means this protocol will simplify the struggle of scientists to isolate neutrophils in terms of steps and expensive material.