Growth, differentiation and the beta-adrenergic signal system of HL-60 cells. Characterization in a medium with insulin as the only added protein.

The purpose of the present investigation was to define experimental conditions for studies on growth, differentiation and the beta-adrenergic signal system of HL-60 cells. The cell medium was completely devoid of added proteins and hormones other than insulin. The HL-60 cell was able to grow and differentiate in this medium. The spontaneous differentiation along the granulocytic pathway after 72 hr, as assessed by the Nitro Blue tetrazolium test, increased by 400% compared to the serum supplemented medium, but the response to 1 microM retinoic acid was equal in the two media. Induction of monocytic differentiation by 0.16 microM phorbol-13-acetate-12-myristate, as determined by surface adherence after 24 hr, was lower in the absence than in the presence of serum. cAMP levels were elevated in response to (-)-isoproterenol. The EC50 was 0.36 +/- 0.01 microM (mean +/- SE, N = 3). The beta-adrenergic ligand 3H-CGP 12177 was specifically bound to 1 single class of binding sites (Kd: 0.15 +/- 0.04 nM, Bmax: 2220 +/- 150, mean +/- SE, N = 3). These data are comparable to our previously reported findings in serum supplemented medium. The present data show that HL-60 cells are able to grow and differentiate in the absence of serum proteins and hormones other than insulin. Under the present experimental conditions, these cells possessed functional beta-adrenergic receptors.