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Cytochemistry and Ultrastructural Morphometry of Cultured HL60 Myeloid Leukemia Cells¹

Departments of Pediatrics and Pathology, University of Texas Health Science Center, San Antonio, Texas 78284-7810 [R. T. P., C. S. G.]; Department of Biochemistry, Emory University School of Medicine, Atlanta, Georgia 30322 [D. T. A., J. M. K.]; and Department of Medicine, University of Alabama, Veterans Administration Medical Center, and Comprehensive Cancer Center, Birmingham, Alabama 35294 [J. C. B.]

Cultured human myeloid leukemia (HL60) cells were characterized using ultrastructural cytochemical methods and differences identified when cells were compared for low (17 to 47), middle (69 to 100), and high (214 to 244) passages or to normal promyelocytes aspirated from bone marrow. Endoplasmic reticulum and transition structures (pre-Golgi compartment) of HL60 cells stained positively for peroxidase using diaminobenzidine but stained sparsely for reducing groups with osmiumzinc iodide. Staining of Golgi elements was relatively indistinct with diaminobenzidine and strong with osmium-zinc iodide, in comparison to freshly harvested promyelocytes which have intense diaminobenzidine and osmium-zinc iodide staining of the pre-Golgi and Golgi compartments. Cytoplasmic polyribosomes were more numerous in middle and high passage cells, whereas dilatation of endoplasmic reticulum was less prominent in these cells. The mean granule size was significantly increased in low passage cells, and staining of peroxidase was more prominent by light and electron microscopy when compared to high passage cells. Cytoplasmic granules demonstrated strong complex carbohydrate staining, indicating a lack of granule maturation in HL60 cells. Terminally differentiated myeloid cells were more frequent in low passage samples, and some neutrophil granule maturation appeared to occur within these cells, whereas all eosinophil granules consistently remained immature with intense complex carbohydrate staining and lack of crystalloid formation. These studies demonstrate significant differences between HL60 cells and normal promyelocytes, and also passage-dependent maturational differences in HL60 cells. These differences should be considered in evaluating parameters of cell growth and maturation and in the biochemical and enzymatic characterization of these cells.

¹ This work was supported in part by Veterans Administration medical research funds and USPHS Grant CA22294 from the National Cancer Institute.

² To whom requests for reprints should be addressed.

Received 12/19/86. Revised 6/18/87. Accepted 6/22/87.