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Hepatocyte growth factor is involved in formation of osteoclast-like cells mediated by clonal stromal cells (MC3T3-G2/PA6)

✍ Scribed by Takuya Sato; Yoshiyuki Hakeda; Yuji Yamaguchi; Hiroshi Mano; Ken-Ichi Tezuka; Kunio Matsumoto; Toshikazu Nakamura; Yoshihisa Mori; Kenji Yoshizawa; Koji Sumitani; Hiroaki Kodama; Masayoshi Kumegawa


Publisher
John Wiley and Sons
Year
1995
Tongue
English
Weight
902 KB
Volume
164
Category
Article
ISSN
0021-9541

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✦ Synopsis


963 (H. K.), lapan KEN-ICHI TEZUKA, K U N I O MATSUMOTO, TOSHIKAZU NAKAMURA,

Osteoclast formation from hemopoietic precursors has been shown to require the support of stromal cells in bone tissue. In this study, we demonstrated that hepatocyte growth factor (HGF) is one of the stromal cell-derived molecules responsible for osteoclast-like cell formation. For our experiments, we used a coculture system for osteoclastic cell formation and activation in which hemopoietic blast cells are cocultured with calvaria-derived stromal MC3T3-G2/PA6 (PA6) cells on dentine slices in the presence of 1,25-dihydroxyvitamin D, [1,25(OH),D,]. Addition of anti-HGF neutralizing IgG to the cocultures inhibited the formation of osteoclastic cells and their dentine-resorbing activity. We detected a single 6.0-kb transcript for HGF in PA6 cells, and also recognized immunoreactive Mr 81,000 and 88,000 forms of HGF in conditioned medium (CM) from PA6 cell cultures, the level of which reached 6 ng/ml. Both the C M and HGF stimulated the proliferation of blast cells synergistically with granulocyte-macrophage colony-stimulating factor, resulting in an increased number of osteoclast precursors that respond to 1,25(OH),D, that are tartrate-resistant acid phosphatase-positive multinucleate cells in stromal cell-free blast cell cultures in plastic wells. The effect of the C M was diminished by the addition of anti-HGF IgG. However, neither the C M nor HGF stimulated the formation of osteoclastic cells and pits on dentine slices in the absence of PA6 cells. These results suggest that although HGF cannot completely replace stromal cells, it is one of the paracrine mediators produced by stromal cells that act on proliferation of osteoclastic cell precursors. o 1995 Wiley-Liss, Inc.