The Netosis Formation of HL-۶۰ Cell Differentiated to Neutrophil-Like Cells by LPS

Background: Neutrophils are the most abundant white blood cells in humans. Recently, a novel strategy called the formation neutrophil extracellular traps (NETs) was described. NETs is a new strategy for pathogen response. This study focused on whether LPS induced NETs release in vitro in the HL۶۰ cell line. Methods: In this study, the HL۶۰ cell line was used for culture and DMSO for induction and differentiation. Flow cytometry was used to evaluate CD۱۱b in the differentiated cells, and the NBT assay was used to evaluate the functionality of the differentiated HL-۶۰ cells. Neutrophil-like cells were incubated with LPS (۲۰۰ ng/ml) for ۴۵ min, followed by incubation for ۲۵ min with ۱۰۰ ng/ml Hoechst ۳۳۳۴۲. Trypan blue as vital staining was used for viability. The statistical significance of the difference between the control and treated groups was evaluated using a one-way ANOVA. Results: Our results showed that ۷۵% NETs was produced by HL-۶۰ differentiated neutrophil cells exposed to ۲۰۰ ng/ml LPS in ۴۵ minues. Conclusion: Consequently, the LPS-induced infection and lethality may occur through various mechanisms. Thus, understanding the molecular mechanisms regulating NET formation in LPS-induced neutrophil-like cells would support the development of new therapeutic methods.

Keywords:

Netosis , Neutrophil , HL-۶۰ , Differentiation , LPS