Using HL-60 Cells to Develop a Model for Studying Neutrophils

Nicholas Sachetta

Abstract:
Neutrophils are the most abundant immune cell in the human body as they make up 50-70% of all white blood cells. The HL-60 cell line is the most commonly used cell line for neutrophil research. Using this cell line, we learned how to grow, split, differentiate, and count HL-60 cells, which has laid the foundation for further experiments in the future, such as inducing NETosis and determining the composition of NETs. HL-60 cells needed to be fed every 2-3 days and split as they approach a concentration of one million cells per milliliter. HL-60 cells can be induced to differentiate into neutrophils when grown in media containing 1.3% of DMSO. We will present data demonstrating that once differentiation has occurred, the viability of dHL-60 cells reduces and the cells stop proliferating, which is similar to neutrophils in the human body, as they only live for a few days. We will also present data suggesting that undifferentiated HL-60 cells have higher metabolic activity than dHL-60 cells, which was unexpected. The use of SDS-PAGE has the potential to show protein differences in differentiated versus undifferentiated HL-60 cells. The study of HL-60 cells can give us more insight into the various functions of neutrophils. Most of these functions are beneficial, but unchecked, some neutrophil functions can lead to autoimmune diseases. Additional research with HL-60 cells will help uncover the mechanisms behind these disease processes.