Abstract
The human leukemia cell line (HL-60) is an alternative to primary neutrophils in research studies. However, because HL-60 cells proliferate in an incompletely differentiated state, they must undergo differentiation before they acquire the functional properties of neutrophils. Here we provide evidence of swarming and chemotaxis in differentiated HL-60 neutrophil-like cells (dHL-60) using precise microfluidic assays. We found that dimethyl sulfoxide differentiated HL-60 cells (DdHL-60) have a larger size, increased length, and lower ability to squeeze through narrow channels compared to primary neutrophils. They migrate through tapered microfluidic channels slower than primary neutrophils, but faster than HL-60s differentiated by other protocols, e.g., using all-trans retinoic acid. We found that dHL-60 can swarm toward zymosan particle clusters, though they display disorganized migratory patterns and produce swarms of smaller size compared to primary neutrophils.
Highlights
The human leukemia cell line (HL-60) is an alternative to primary neutrophils in research studies
We found that 65%, 39%, and 1% of human neutrophils, differentiated HL-60 cells (DdHL-60), and AdHL-60 migrated in fMLP gradients, respectively (Fig. 2b)
The swarm size for similar targets is smaller and the trajectories of cells joining the swarms appear more disorganized for differentiated HL-60 cells compared to primary neutrophils
Summary
The human leukemia cell line (HL-60) is an alternative to primary neutrophils in research studies. We provide evidence of swarming and chemotaxis in differentiated HL-60 neutrophil-like cells (dHL-60) using precise microfluidic assays. We found that dimethyl sulfoxide differentiated HL-60 cells (DdHL-60) have a larger size, increased length, and lower ability to squeeze through narrow channels compared to primary neutrophils. They migrate through tapered microfluidic channels slower than primary neutrophils, but faster than HL-60s differentiated by other protocols, e.g., using all-trans retinoic acid. We employed microfluidic migration assays and a micropatterning array to compare the swarming and migratory ability of HL-60 cells after they were differentiated using DMSO (DdHL-60), all-trans-retinoic acid (AdHL-60) and nutridoma supplemented DMSO (nDdHL-60). Our study suggests that differences in migration, deformability, and persistence have to be taken into account when employing differentiated HL-60 cells as models for studying primary neutrophil functions
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