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Retinoic Acid-Induced HL-60 Cell Differentiation Is Augmented by Copper Supplementation^1,2,3,4,

Food Science and Human Nutrition Department, University of Florida, Gainesville, FL 32611

The objective of this study was to determine how copper influences the ability of HL-60 cells to differentiate into cells of the granulocytic lineage. We hypothesized that granulopoiesis requires copper because copper-deficient humans become neutropenic. Differentiation of HL-60 cells along the granulocytic lineage with retinoic acid was enhanced by copper. The results showed a greater number of cells were more differentiated when copper was added to the medium for 96 h. The respiratory burst activity of retinoic acid-induced cells was increased by copper supplementation, but intracellular superoxide anion generation was not affected. Supplementation with copper resulted in more cell-associated copper in both noninduced and induced cells; however, the induced cells accumulated three times more copper than the noninduced cells. Even though the amount of copper associated with retinoic acid-treated cells was greater than in untreated cells, the activity of a copper-requiring enzyme, copper/zinc superoxide dismutase, was significantly lower. Copper supplementation increased the activity of this enzyme in both retinoic acid-treated and untreated cells. Cytochrome c oxidase activity was not affected by retinoic acid treatment or by copper supplementation. Copper seems to play a specific role during the early stages of granulocyte differentiation.

KEY WORDS: • copper • HL-60 cells • copper status • respiratory burst • differentiation

¹ Presented in part at the 75th Annual Meeting of the Federation of American Societies for Experimental Biology, April 1991, Atlanta, GA [Bae, B. & Percival, S. S. (1992) Copper's role in HL-60 cell differentiation. FASEB J. 5: A1076 (abs. 4094).

² Supported in part by NIH grant DK45813 and the International Life Sciences Institute/Nutrition Foundation Future Leader Award, and funds provided by the Center for Nutritional Sciences, University of Florida.

³ Florida Agriculture Experiment Station Journal Series number R-02724.

⁴ The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked "advertisement" in accordance with 18 USC section 1734 solely to indicate this fact.

⁵ To whom correspondence should be addressed.

Manuscript received 13 October 1992. Revision accepted 26 January 1993.