Functional changes in human leukemic cell line HL-60. A model for myeloid differentiation

Abstract

Polar solvents induce terminal differentiation in the human promyelocytic leukemia cell line HL-60 . The present studies describe the functional changes that accompany the morphologic progression from promyelocytes to bands and polymorphonuclear leukocytes (PMN) over 9 d of culture in 1 .3% dimethylsulfoxide (DM SO). As the HL-60 cells mature, the rate of 02 production increases 18-fold, with a progressive shortening of the lag time required for activation . Hexosemonophosphate shunt activity rises concomitantly . Ingestion of paraffin oil droplets opsonized with complement or Ig increases 10-fold over 9 d in DMSO. Latex ingestion per cell by each morphologic type does not change significantly, but total latex ingestion by groups of cells increases with the rise in the proportion of mature cells with greater ingestion capacities . Degranulation, as measured by release of R-glucuronidase, lysozyme, and peroxidase, reaches maximum after 3-6 d in DMSO, then declines. HL-60 cells contain no detectable lactoferrin, suggesting that their secondary granules are absent or defective . However, they kill staphylococci by day 6 in DMSO. Morphologically immature cells (days 1-3 in DMSO) are capable of 02 generation, hexosemonophosphate shunt activity, ingestion, degranulation, and bacterial killing. Maximal performance of each function by cells incubated in DMSO for longer periods of time is 50-100% that of normal PMN . DMSO-induced differentiation of HL-60 cells is a promising model for myeloid development .KEY WORDS phagocytosis " superoxide granulocyte -lysosome -lactoferrin A human promyelocytic leukemia cell line, HL-60, was recently established from the peripheral blood of a patient with acute promyelocytic leukemia (9) . It has maintained continuous growth in J . CELL. BIOLOGY © The Rockefeller University Press -0021-9525/79/08/0315/08 $1 .00 Volume 82 August 1979 315-322 suspension culture in the absence of added conditioned medium or colony-stimulating factor for over 18 mo and is tumorigenic in athymic nude mice (10). The majority of HL-60 cells are promyelocytic in morphology and histochemistry, but 4-15% of them show morphologic characteristics of more mature myeloid cells: myelocytes, meta-315 on

Optimization of a myeloid cell transfusion strategy for infected neutropenic hosts

“…The HL-60 cell line was originally obtained by leukapheresis of peripheral blood cells from a patient with acute myelogenous leukemia [25] and has been maintained in culture without genetic manipulation since that time [package insert, American Type Culture Collection (ATCC), Manassas, VA, USA]. A major advantage of HL-60 cells is that they can be activated to differentiate toward a mature neutrophil phenotype [26]. Specifically, when activated for 7 days in the presence of retinoic acid (RA) and DMSO, we found a significant improvement in HL-60 cell phagocytosis and killing of the pathogenic fungus, Candida albicans in vitro [24].…”

Appearance of an arachidonic acid 15‐lipoxygenase pathway upon differentiation of the human promyelocytic cell‐line HL‐60

“…Formation of the pair of C-g epimeric isomers of 8, I5-DHETE can be observed upon incubation of IS-HPETE with various heme-proteins [12]. Thus, in the present study the contribution of ~eme-proteins to the formation of 8,115DHETE cannot be ruled out since protein synthesis increases during differentiation [4] of these cells.…”

“…Differentiated HL-60 cells also develop a capacity to initiate an oxidative burst [4], This ability has been shown to parallel increments in NADPHoxidase activity, Also, increased capacity for O;-production parallels increments in the levels of cytochrome b in these ceils f4]. Formation of the pair of C-g epimeric isomers of 8, I5-DHETE can be observed upon incubation of IS-HPETE with various heme-proteins [12].…”

Developmental Expression of Insulin Receptor Substrate-2 During Dimethylsulfoxide-Induced Differentiation of Human HL-60 Cells

“…HL-60 cells differentiated in the presence of DMSO are well known to display several functional characteristics of granulocytes, including superoxide anion production, phagocytosis, and chemotaxis (38,39). For example, these cells are 90% positive for superoxide anion production after 96 h in 1.2% DMSO (40).…”