Factors responsible for variable reported lineages of HL-60 cells induced to mature with butyric acid.
HL-60 is a multipotential human leukemia cell line widely used as an in vitro model to investigate myeloid differentiation. A variety of compounds can reproducibly induce these cells to differentiate towards specific lineages. However, under what appear to be similar experimental conditions, various laboratories have reported either neutrophilic differentiation or monocytic differentiation after butyric acid induction. We investigated different hypotheses to explain these dissimilar findings. First, the potential role of variable 1,25-dihydroxyvitamin D3 (VD3) concentrations in commercial fetal calf serum was assessed. Second, possible differences between laboratories inherent to the HL-60 cells themselves were explored. Lineage was assessed by morphology, histochemistry (nonspecific esterase), and neutrophil-specific (CD15) and monocyte-specific (MO2) surface antigens. We found that increasing concentrations of VD3 spanning the range reported in commercial fetal calf serum (25 to 155 pg/ml) act in synergy with butyric acid to result in higher monocyte/neutrophil ratios at the higher VD3 concentrations. Different lots of serum led to monocyte/neutrophil ratios in proportion to their VD3 concentrations. Starting with HL-60 cells obtained from different laboratories, several single-cell clones were derived which yielded either high percentages of monocytes, high percentages of neutrophils, or intermediate mixes of both cell types after induction with butyric acid. We conclude that the wide variation of VD3 concentration found in different lots of commercial fetal calf serum and intrinsic differences in HL-60 cells are two identifiable factors that can explain the discrepancies in lineage observed by different investigators after butyric acid induction of HL-60 cells.
Hoessly
M C
MC
Hematology/Oncology Section, University of Pennsylvania School of Medicine, Philadelphia 19104-4283.
Rossi
R M
RM
Fischkoff
S A
SA
D000943:Antigens, Differentiation / Q000032:analysis; D002087:Butyrates / Q000494:pharmacology*; D020148:Butyric Acid; D002117:Calcitriol / Q000494:pharmacology; D043182:Carboxylesterase; D002265:Carboxylic Ester Hydrolases / Q000378:metabolism; D002454:Cell Differentiation / Q000187:drug effects*; D002469:Cell Separation; D006801:Humans; D015470:Leukemia, Myeloid, Acute / Q000473:pathology*; D016256:Lewis X Antigen; D008264:Macrophages / Q000166:cytology; D009000:Monocytes / Q000166:cytology; D009504:Neutrophils / Q000166:cytology*; D014407:Tumor Cells, Cultured
D000943:Antigens, Differentiation; D002087:Butyrates; D016256:Lewis X Antigen; D020148:Butyric Acid; D002265:Carboxylic Ester Hydrolases; D043182:Carboxylesterase; D002117:Calcitriol