Development and characterization of a human marrow stromal cell line that enhances osteoclast-like cell formation.
We established a human bone marrow stromal cell line (Saka) by infecting marrow adherent cells from semisolid marrow cultures with a recombinant simian virus-40 (SV40) virus. The cells expressed SV40 large tumor antigen, had a fibroblast-like shape, and expressed fibronectin and vimentin. They did not contain detectable alkaline phosphatase activity; express myeloid, lymphoid, or factor VIII-associated antigens; or develop adipocyte-like characteristics with dexamethasone treatment. Polymerase chain reaction analysis of Saka cell RNA detected expression of messenger RNAs for interleukin-6 (IL-6), IL-1 beta, granulocyte-macrophage colony-stimulating factor, macrophage colony-stimulating factor, stem cell factor, and the 1,25-dihydroxyvitamin D3 receptor. Coculture of Saka cells with human marrow mononuclear cells enhanced formation of osteoclast-like multinucleated cells (MNC) in long term human bone marrow cultures. These MNC expressed calcitonin receptors and formed resorption lacunae on dentine. In contrast, coculture of marrow mononuclear cells with other SV40-transformed human marrow stromal cell lines did not increase MNC formation. Conditioned medium from Saka cells or coculture of bone marrow and Saka cells separated by a Millipore membrane did not enhance MNC formation. Addition of a neutralizing antibody to IL-6 or IL-1 beta blocked the effects of Saka cells on MNC formation. These results suggest that marrow stromal cells enhance osteoclast formation in part through direct cell to cell contact and production of IL-6 and/or IL-1 beta.
Takahashi
S
S
Audie Murphy Veterans Administration Hospital, San Antonio, Texas 78284.
Reddy
S V
SV
Dallas
M
M
Devlin
R
R
Chou
J Y
JY
Roodman
G D
GD
D001483:Base Sequence; D001854:Bone Marrow Cells; D002460:Cell Line; D002461:Cell Line, Transformed; D005347:Fibroblasts / Q000166:cytology; D005353:Fibronectins / Q000032:analysis; D016178:Granulocyte-Macrophage Colony-Stimulating Factor / Q000235:genetics; D016298:Hematopoietic Cell Growth Factors / Q000235:genetics; D006801:Humans; D007375:Interleukin-1 / Q000235:genetics; D015850:Interleukin-6 / Q000235:genetics; D016173:Macrophage Colony-Stimulating Factor / Q000235:genetics; D008969:Molecular Sequence Data; D010010:Osteoclasts / Q000166:cytology*; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis / Q000378:metabolism; D018167:Receptors, Calcitriol / Q000235:genetics; D013539:Simian virus 40; D019089:Stem Cell Factor; D017154:Stromal Cells / Q000502:physiology*; D014746:Vimentin / Q000032:analysis
D005353:Fibronectins; D016298:Hematopoietic Cell Growth Factors; D007375:Interleukin-1; D015850:Interleukin-6; D012333:RNA, Messenger; D018167:Receptors, Calcitriol; D019089:Stem Cell Factor; D014746:Vimentin; D016173:Macrophage Colony-Stimulating Factor; D016178:Granulocyte-Macrophage Colony-Stimulating Factor