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Effect of fat-cell conditioned-medium on differentiation of MC3T3-E1

DU Yu,Guo Feng-jing,XU Fei,et al.Department of Orthopedics,Tongji Hospital,Tongji Medical College,Huazhong University of Science and Technology,Wuhan 430030,China  
Objective To investgate the influence of fat-cell conditioned-medium(FCCM) on the differentiation of MC3T3-E1.Methods FCCM and control medium(CM) were prepared to culture MC3T2-E1 cells.The expression of Runt-related transcription factor 2(Runx2),alkaline phosphatase(ALP),and bone gamma-carboxyglutamic-acid-containing protein(BGP) was detected by using real-time PCR and Western blotting.ALP activity was determined by pnitro phenyl phosphate assay.Calcium nodules were examined by using Van Gieson and alizarin red method.Results The expression of Runx2,ALP and BGP,and activity of ALP in FCCM group were reduced as compared with CM group.ALP staining and Alizarin red staining in FCCM group was weaker than CM group.Conclusion FCCM can down-regulated the osteogenic differentiation of MC3T3-E1.
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