Osteoinduction in human fat derived stem cells by recombinant human bone morphogenetic protein-2 produced in Escherichia coli

摘要

Bioactive recombinant human bonemorphogenetic protein-2 (rhBMP-2) was obtainedusing Escherichia coli pET-25b expression system:55 mg purified rhBMP-2 were achieved per g cell drywt, with up to 95% purity. In murine C2C12 cell line,rhBMP-2 induced an increase in the transcription ofSmads and of osteogenic markers Runx2/Cbfa1 andOsterix, measured by semi-quantitative RT-PCR.Bioassays performed in human fat-derived stem cellsshowed an increased activity of the early osteogenicmarker, alkaline phosphatase, and the absence ofcytotoxicity.