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Induction of Monocyte-Macrophage Differentia of Human Hematopoietic Cells (CM-S) by Phorbol Esters

Institute of Comparative Anatomy, University of Rome, [G. M.]; Institute of Genetics, University of Naples, Naples, [M. L.]; Institute of Pediatrics G. Gaslini, Genoa [P. C.-F.]; First Institute of Clinical Medicine, University of Milan, Milan, [G. L. D.]; and Institute of Cell Biology, C. N. R., Via G. D. Romagnosi 18A, 00196 Rome [E. V., R. R.], Italy

A human diploid hematopoietic cell line, CM-S, has been established from the bone marrow of a patient with congenital hypoplastic (Diamond-Blackfan) anemia. The cells grow in suspension in an almost undifferentiated form but express some enzyme markers of monocytic and granulocytic cells. Upon treatment with the tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA)² (10^-7 M concentrations or higher), the cells become rapidly adherent to glass or plastic surfaces and show, by transmission electron microscopy, an increase in the rough endoplasmic reticulum, an apparent increase in the number and size of swollen mitochondria, and numerous granules, rarely seen in the absence of TPA. Fc and C3 receptors increase, and the cells become capable of phagocytosis and immunoerythrophagocytosis. Cytochemical analysis of the treated cells shows an increase in cells positive for a variety of enzyme markers usually associated with monocytic cells [e.g., acid phosphatase, ß-galactosidase, -naphthyl acetate esterase (mostly fluoride sensitive), cytochrome oxidase, 7ga-naphthol acetate esterase] and a corresponding decrease in cells positive for granulocytic markers (e.g., myeloperoxidase, Sudan black, and naphthol ASD chloroacetate esterase). Significant changes are also seen in the expression of ß₂-microglobuln, HLA-ABC, and la-like antigens. No evidence was found, either in induced or in uninduced cells, for B- or T-lymphocyte markers. TPA affects proliferation of CM-S cells. Cell differneitation is related to the structure of the phorobol diester. CM-S cells upon induction with TPA thus possess the capacity to differentiate and acquire several morphological and functional characteristics of monocyte-macrophages.

¹ To whom requests for reprints should be addressed.

Received 9/25/81. Accepted 6/10/82.

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