title: Dimethylsulfoxide exposure modulates HL-60 cell rolling interactions.
abstract: Human leukaemic HL-60 cells are widely used for studying interactions involving adhesion molecules [e.g. P-selectin and PSGL-1 (P-selectin glycoprotein ligand-1)] since their rolling behaviour has been shown to mimic the dynamics of leucocyte rolling in vitro. HL-60 cells are neutrophilic promyelocytes that can undergo granulocytic differentiation upon exposure to compounds such as DMSO (dimethylsulfoxide). Using a parallel plate flow chamber functionalized with recombinant P-selectin-Fc chimaera, undifferentiated and DMSO-induced (48, 72 and 96 h) HL-60 cells were assayed for rolling behaviour. We found that depending on P-selectin incubation concentration, undifferentiated cells incurred up to a 6-fold increase in rolling velocity while subjected to an approximately 10-fold increase in biologically relevant shear stress. HL-60 cells exposed to DMSO for up to 72 h incurred up to a 3-fold increase in rolling velocity over the same shear stress range. Significantly, cells exposed for up to 96 h incurred up to a 9-fold decrease in rolling velocity, compared with undifferentiated HL-60 cells. Although cell surface and nuclear morphological changes were evident upon exposure to DMSO, flow cytometric analysis revealed that PSGL-1 expression was unchanged, irrespective of treatment duration. The results suggest that DMSO-treated HL-60 cells may be problematic as a substitute for neutrophils for trafficking studies during advanced stages of the LAC (leucocyte adhesion cascade). We suggest that remodelling of the cell surface during differentiation may affect rolling behaviour and that DMSO-treated HL-60 cells would behave differently from the normal leucocytes during inflammatory response in vivo.
authors.affiliation: Department of Mechanical Engineering, Rochester Institute of Technology, 76 Lomb Memorial Drive, Rochester, NY, USA. David_Gee@mail.RIT.edu
authors.forename: David J
authors.initials: DJ
authors.lastname: Gee
authors.forename: L Kate
authors.initials: LK
authors.lastname: Wright
authors.forename: Jonathan
authors.initials: J
authors.lastname: Zimmermann
authors.forename: Kayla
authors.initials: K
authors.lastname: Cole
authors.forename: Karen
authors.initials: K
authors.lastname: Soule
authors.forename: Michelle
authors.initials: M
authors.lastname: Ubowski
mesh_terms: D002448:Cell Adhesion; D002467:Cell Nucleus / Q000187:drug effects; D048430:Cell Shape / Q000187:drug effects; D003593:Cytoplasm / Q000187:drug effects; D004121:Dimethyl Sulfoxide / Q000494:pharmacology*; D018922:HL-60 Cells; D006801:Humans; D055767:Immobilized Proteins / Q000378:metabolism; D007141:Immunoglobulin Fc Fragments / Q000378:metabolism; D036904:Leukocyte Rolling / Q000187:drug effects*; D008562:Membrane Glycoproteins / Q000378:metabolism; D019007:P-Selectin / Q000378:metabolism; D011993:Recombinant Fusion Proteins / Q000378:metabolism
chemical_list: D055767:Immobilized Proteins; D007141:Immunoglobulin Fc Fragments; D008562:Membrane Glycoproteins; D019007:P-Selectin; C084882:P-selectin ligand protein; D011993:Recombinant Fusion Proteins; D004121:Dimethyl Sulfoxide