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Generation of Avian Cells Resembling Osteoclasts from Mononuclear Phagocytes^*

JOSE I. ALVAREZ, STEVEN L. TEITELBAUM, HARRY C. BLAIR, EDWARD M. GREENFIELD, NICHOLAS A. ATHANASOU and F. PATRICK ROSS

Department of Pathology and Laboratory Medicine (J.I.A., S.L.T., H.C.B., E.M.G., F.P.R.), Jewish Hospital at Washington University Medical Center St. Louis, Missouri 63110
Centro Nacional de Enfermedades Reumaticas (J.I.A.), Hospital Universitario de Caracas Caracas 1010, Venezuela
Nuffield Department of Pathology (N.A.A.), University of Oxford, John Radcliffe Hospital Oxford 0X3 9DU, United Kingdom

Address all correspondence and requests for reprints to: Dr. F. Patrick Ross, Department of Pathology and Laboratory Medicine, Jewish Hospital at Washington University Medical Center, 216 South Kingshighway, St. Louis, Missouri 63110.

Abstract

Several lines of indirect evidence suggest that a monocyte family precursor gives rise to the osteoclast, although this hypothesis is controversial. Starting with a uniform population of nonspecific esterase positive, tartrate-sensitive, acid phosphatase-producing, mannose receptor-bearing mononuclear cells, prepared from dispersed marrow of calcium-deprived laying hens by cell density separation and selective cellular adherence, we generated multinucleated cells in vitro. When cultured with devitalized bone, these cells show, by electron microscopy, the characteristic osteoclast morphology in that they are mitochondria- rich, multinucleated, and, most importantly, develop characteristic ruffled membranes at the matrix attachment site. Moreover, as documented by scanning electron microscopy, these cells pit bone slices in a manner identical to freshly isolated osteoclasts. In addition, isoenzymes of acid phosphatase from generated osteoclasts, separated by 7.5% polyacrylamide gel electrophoresis at pH 4, are identical to those of mature osteoclasts in migration pattern and tartrate resistance, although the precursor cells from which the osteoclasts are generated produce an entirely different isoenzyme, which is tartrate-sensitive and migrates less rapidly at pH 4. The fused cells also exhibit a cAMP response to prostaglandin E₂. Therefore, osteoclast-like cells can be derived by in vitro culture of a marrow-derived monocyte cell population. (Endocrinology 128: 2324–2335, 1991)

Footnotes

^* This work was supported in part by Grant AR-32788 from the NIH and by a grant from Saint Louis Shriners Hospital.

Recipient of a grant from the Fundacion Gran Mariscal de Ayacucho, Caracas, Venezuela.

NASA Research Associate NAGW-70.

Received December 21, 1990.

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