Application of DUSP6 inhibitor BCI in preparation of osteoporosis drugs
A technology of osteoporosis and inhibitors, applied in the field of medicine, can solve the problems of unclear effects on osteoclast differentiation-related diseases
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[0029] Cell culture and osteoclast differentiation
[0030] Mouse macrophages RAW264.7 were derived from ATCC cell bank (Shanghai, China) and cultured in DMEM (10% fetal bovine serum and 1% P / S). Bone marrow mononuclear macrophages (BMMs) were isolated from mouse femur and tibia, and cultured in α-MEM (10% fetal bovine serum, 1% penicillin / streptomycin and 50ng / mL M-CSF) for 48 hours. All cells in 5% CO 2Incubate at 37°C. 5 hours before osteoclast induction, DUSP6-specific small interfering RNA (siRNA) oligonucleotide (40 nM; 5'-CGAUGCUUACGACAUUGUUUAATTUUAACAAUGUCGUAAGCAUCGTT-3') (Shanghai Shengong) was transfected by Lipofectamine 3000 (Invitrogen). Both cell types were differentiated using M-CSF (50 ng / mL) and RANKL (100 ng / mL). The degree of osteoclast differentiation of RAW264.7 cells and BMMs was determined on the 3rd and 5th day of induction, respectively. MC3T3-E1 cells (pre-osteogenic cell line) were grown in α-MEM (10% fetal calf serum and 1% penicillin / streptomyc...
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