Vol. 172, No. 1, 1990 October 15, 1990 BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS Pages 295-299 THE NON-OSTEOGENIC MOUSE PLURIPOTENT CELL LINE, C3H10T1/2, IS INDUCED TO DIFFERENTIATE INTO OSTEOBLASTIC CELLS BY RECOMBINANT HUMAN BONE MORPHOGENETIC PROTEIN-2 Takenobu Katagiri 1,2, Akira Yamaguchi 3, Tohru Ikeda 3, Shusaku Yoshiki 3, John M. Wozney 5, Vicki Rosen 5, Elizabeth A. Wang 5, Haruo Tanaka 2, Satoshi Omura 4, and Tatsuo Suda 1# Department of IBiochemistry and 3Oral Pathology, School of Dentistry, Showa University, 1-5-8 Hatanodai, Shinagawa-ku, Tokyo t42, Japan 2School of Pharmaceutical Sciences, Kitasato University and 4The Kitasato Institute, 5-9-1 Shirokane, Minato-ku, Tokyo 108, Japan 5Genetics Institute Inc., Cambridge, MA Received August 29, 1990 SUMMARY: The possibility that the non-osteogenic mouse pluripotent cell line, C3H10T1/2 (10T1/2), could be induced to differentiate into osteogenic cells by various hormones and cytokines was examined in vitro. Of a number of agents tested, recombinant human bone morphogenetic protein-2 (rhBMP-2) and retinoic acid induced alkaline phosphatase (ALP) activity in 10T1/2 cells, rhBMP-2 also induced mRNA expression of ALP in the cells. Dexamethasone, 1et,25-dihydroxyvitamin D3, transforming growth factor-Ill and insulin-like growth factor-I did not stimulate ALP activity. Treatment with rhBMP-2 greatly induced cAMP production in response to parathyroid hormone in 10T1/2 cells. No ALP activity was induced in NIH3T3 fibroblasts treated with rhBMP-2 or retinoic acid. These results indicate that 10T1/2 ceils have a potential to differentiate into osteogenic cells under the control of BMP-2. o 1990 ~cade~ucPress, Tnc. It is believed that osteoblasts are developed from undifferentiated mesenchymal progenitor cells, which can also differentiate into other specialized connective tissue cells such as chondrocytes, muscle cells and adipocytes (1,2). Several osteoblast-like cells have been used to study the mechanism of osteoblast differentiation in in vitro systems. But, such osteoblast-like cells are inappropriate to explore the events occurring in the early stage of osteoblast differentiation from undifferentiated mesenchymal progenitor cells. To understand precisely the mechanism of osteoblast differentiation, an appropriate in vitro system is required, which reflects the early stage of differentiation from uncommitted mesenchymal cells into osteoblasts. The mouse fibroblastic cell line, C3H10T1/2 clone 8 (10T1/2) established from an early mouse embryo (3), has the pluripotent activity to differentiate into #To whom correspondence should be addressed. 295 0006-291X/90 $1,50 Copyright © 1990 by Academic Press, Inc. All rights of reproduction in any form reserved.