Regulating effect of bone morphogenetic protein-2,-4, and-6 on chondrocyte formation from adipose derived stem cells

Abstract

Objective To observe the influence of cytokines of bone morphogenetic protein (BMP)-2,-4,and-6 on the process of chondrocyte formation from rabbit adipose derived stem cells (ADSCs).Methods The surface antigens CD34,CD45,CD44 and CD29 of ADSCs were identified by flow cytometry,and ADSCs were cultured for 21 days by using the chondrocyte inducing conditional media serving as the culture media containing transforming growth factor (TGF)-β3,and respectively 100 μg/L BMP-2,BMP-4 or BMP-6 to experiment groups.Acidic glycosaminoglycans were detected by alcian blue staining.The mRNA and protein expression levels of aggrecan (ACAN),type Ⅱ collagen (COL2A1) and sex determining region Y-box 9 (SOX-9) were analyzed by real-time quantitative polymerase chain reaction (Real-time PCR),and Western blotting,respectively.Results Only cell proliferation is realized in BMP-4 group,while cell proteoglycan expression levels were more obviously raised in BMP-6 group than in BMP-2 group.The mRNA expression levels of ACAN,COL2A1,and SOX-9 in BMP-6 group were separately 1.7,1.3 and 1.0 times higher than those in BMP-2 group.And protein expression levels of ACAN,COL2A1,and SOX-9 in BMP-6 group were 1.4,1.3 and 0.9 times higher than those in BMP-2 group.Conclusion The combination of TGF-β3 and BMP-6 can promote higher efficiency of differentiation of ADSCs into chondrocytes than BMP-2 and BMP-4. Key words: Adipose derived stem cells; Bone morphogenetic protein ; Chondrocytes ; Translant growth factor-3; Tissue engineering