Biochemical and Biophysical Research Communications
Regular ArticleInduction of Mononuclear Precursor Cells with Osteoclastic Phenotypes in a Rat Bone Marrow Culture System Depleted of Stromal Cells
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Extremely High Expression of Antisense RNA for Wilms' Tumor 1 in Active Osteoclasts: Suppression of Wilms' Tumor 1 Protein Expression during Osteoclastogenesis
2016, American Journal of PathologyCitation Excerpt :After 3 days, the cells were stained for tartrate-resistant acid phosphatase, and tartrate-resistant acid phosphatase–positive multinucleated cells were counted. Osteoclasts were also generated from rat bone marrow cultures as described previously.28,29 Bone marrow cells were obtained from tibia and femurs of male SD rats and cultured for 4 days in 24-well plates (1 × 106 cells per well) in the presence of 10−8 mol/L 1α,25-dihydroxyvitamin D3, 10 ng/mL RANKL, and 10% (v/v) heat-treated conditioned medium of rat osteoblastic cell line ROS17/2.8 (htROSCM).
Identification of two-pore channel 2 as a novel regulator of osteoclastogenesis
2012, Journal of Biological ChemistryCitation Excerpt :Mouse bone marrow cells were obtained from the tibia and femur of mice (C57BL/6, 4-week-old males; Charles River). Bone marrow stromal cells were depleted from mouse bone marrow macrophage (BMM) cells using the Sephadex G-10 (GE Healthcare) column as described previously (18–20). They were cultured in α-MEM (Invitrogen) supplemented with 10% fetal bovine serum (FBS) and 1% penicillin and streptomycin (PS) with 20 ng/ml macrophage colony-stimulating factor (M-CSF; R&D Systems).
Regulation of osteoclastogenesis by ganoderic acid DM isolated from Ganoderma lucidum
2009, European Journal of PharmacologyTwo histone deacetylase inhibitors, trichostatin A and sodium butyrate, suppress differentiation into osteoclasts but not into macrophages
2003, BloodCitation Excerpt :Rabbit polyclonal antibodies against NF-κB p65 and phosphorylated p38 (Tyr 182)–R and mouse monoclonal antibody against actin (C-2) were purchased from Santa Cruz Biotechnology (Santa Cruz, CA). Cultures of rat bone marrow cells were carried out as described by Kukita et al.34-35 Briefly, bone marrow cells were isolated from tibias and femurs of rats. For the formation of MNCs, whole bone marrow cells were cultured in 24-well culture plates (Falcon, Lincoln Park, NJ) for 5 days in α-MEM containing 15% FCS in the presence of 10−8 M 1α,25(OH)2D3 and 10% (vol/vol) heat-treated conditioned medium derived from rat osteoblastic cell line ROS17/2.8 (htROSCM), as described by Kukita et al.34 For the formation of POCs, stroma-free bone marrow cells (2 × 106 cells/mL) were cultured in 96-well culture plates (Falcon) for 4 days in α-MEM containing 15% FCS in the presence of 10−8 M 1α,25(OH)2D3and 10% (vol/vol) heat-treated conditioned medium and TNF-α (10 ng/mL) as described previously.33,35