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Production of Granulocyte-Macrophage (GM-CSF) and Granulocyte Colony-Stimulating Factor (G-CSF) by Rat Clonal Osteoblastic Cell Population CRP 10/30 and the Immortalized Cell Line IRC10/30-mycl Stimulated by Tumor Necrosis Factor ^*

Department of Pathophysiology, University of Berne CH-3010 Berne, Switzerland

Abstract

Previously, we have shown that primary cultures of murine calvarial cells produce both granulocyte macrophage (GM) and granulocyte (G) colony stimulating factor (CSF). Because of the heterogeneity of cell types in these cultures the osseous origin of these cytokines was not certain. Thus a nontransformed rat clonal osteoblastic cell population CRP 10/30 and the immortalized cell line IRClO/30-mycl derived from it, which both express the osteoblastic phenotype, were now investigated. Both produced hemopoietic growth activity after treatment with recombinant murine tumor necrosis factor a. This activity eluted from diethylaminoethyl Sephacel at 0.2-0.3 M NaCl, and migrated on Sephacryl S-200 at a mol wt of around 30 k, as described for murine GM- and G-CSF. On a Phenyl Sepharose CL-4B column, it was separated into two peaks appearing at position where GM (peak I) and G-CSF (peak II) are expected to be eluted. Antisera against GM-CSF inhibited the activity of peak I. In the colony assay in semisolid medium, peak I induced colonies of the GM-type and peak II of the Gtype. These data indicate that cloned osteoblasts produce GMand G-CSF. Through CSF production, osteoblasts might regulate osteoclast formation, influence hemopoiesis and/or participate in local inflammatory reactions of bone. (Endocrinology 128: 661–667, 1991)

Footnotes

^* This work has been supported by the Swiss National Science Foundation Grant 3.894-0.88 SR).

Received June 21, 1990.

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