methylcellulose and glycyl-arginyl-glycyl-aspartyl-serine

Identification of receptor activator of nuclear factor-kappaB (RANK) and RANK-ligand (RANKL) has provided new insights into the osteoclast differentiation pathway. Osteoclast precursor cells were isolated using monoclonal antibodies against c-Fms and RANK, and the effect of adherence on the in vitro differentiation and proliferation of these cells was examined in 2 different types of stromal-cell-free culture systems: a semisolid culture medium (a nonadherent system) and a liquid culture medium (an adherent system). Osteoclast precursor cells were not able to differentiate into mature osteoclasts efficiently in the semisolid culture system. Trimerized RANKL enhanced osteoclast differentiation in semisolid cultures, but not to the extent seen when cells were allowed to adhere to plastic. Initial precursor cells were capable of differentiating into macrophages or osteoclasts. Once these cells were transferred to adherent conditions, striking differentiation was induced. Multinuclear cells were observed even after they had displayed phagocytic activity, which suggests that cell adhesion plays an important role in the differentiation of osteoclast precursor cells. Integrins, especially the arginine-glycine-aspartic acid (RGD)-recognizing integrins alpha(v) and beta(3), were needed for osteoclast-committed precursor cells to proliferate in order to form multinuclear osteoclasts, and the increase in cell density affected the formation of multinuclear cells. A model of osteoclast differentiation with 2 stages of precursor development is proposed: (1) a first stage, in which precursor cells are bipotential and capable of anchorage-independent growth, and (2) a second stage, in which the further proliferation and differentiation of osteoclast-committed precursor cells is anchorage-dependent. (Blood. 2000;96:4335-4343)

Topics: Animals; Antibodies, Monoclonal; Antigens, CD; Carrier Proteins; Cell Adhesion; Cell Culture Techniques; Cell Differentiation; Cell Division; Cells, Cultured; Culture Media; Female; Giant Cells; Glycoproteins; Hematopoietic Stem Cells; Integrin alphaV; Integrin beta3; Leucine Zippers; Macrophage Colony-Stimulating Factor; Macrophages; Membrane Glycoproteins; Methylcellulose; Mice; Mice, Inbred C57BL; Models, Biological; Oligopeptides; Osteoclasts; Osteoprotegerin; Plastics; Platelet Membrane Glycoproteins; RANK Ligand; Receptor Activator of Nuclear Factor-kappa B; Receptor, Macrophage Colony-Stimulating Factor; Receptors, Cytoplasmic and Nuclear; Receptors, Tumor Necrosis Factor