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Expression, purification and osteogenic bioactivity of recombinant human BMP-4, -9, -10, -11 and -14

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Resumo:Bone morphogenetic proteins (BMPs) are cytokines from the TGF-b superfamily, with important roles during embryonic development and in the induction of bone and cartilage tissue differentiation in the adult body. In this contribution, we report the expression of recombinant human BMP-4, BMP-9, BMP-10, BMP-11 (or growth differentiation factor- 11, GDF-11) and BMP-14 (GDF-5), using Escherichia coli pET-25b vector. BMPs were overexpressed, purified by affinity his-tag chromatography and shown to induce the expression of early markers of bone differentiation (e.g. smad-1, smad-5, runx2/cbfa1, dlx5, osterix, osteopontin, bone sialoprotein and alkaline phosphatase) in C2C12 cells and in human adipose stem cells. The described approach is a promising method for producing large amounts of different recombinant BMPs that show potential for novel biomedical applications.
Autores principais:Bessa, Paulo C.
Outros Autores:Cerqueira, M. T.; Rada, Tommaso; Gomes, Manuela E.; Neves, N. M.; Nobre, Alexandra; Reis, R. L.; Casal, Margarida
Assunto:Bone morphogenetic protein-4 Bone morphogenetic protem-9 Bone morphogenetic protein-10 Growth differentiation factor-11 Growth differentiation factor-5 C2C12 Bone morphogenetic protein-9
Ano:2009
País:Portugal
Tipo de documento:artigo
Tipo de acesso:acesso aberto
Instituição associada:Universidade do Minho
Idioma:inglês
Origem:RepositóriUM - Universidade do Minho
Descrição
Resumo:Bone morphogenetic proteins (BMPs) are cytokines from the TGF-b superfamily, with important roles during embryonic development and in the induction of bone and cartilage tissue differentiation in the adult body. In this contribution, we report the expression of recombinant human BMP-4, BMP-9, BMP-10, BMP-11 (or growth differentiation factor- 11, GDF-11) and BMP-14 (GDF-5), using Escherichia coli pET-25b vector. BMPs were overexpressed, purified by affinity his-tag chromatography and shown to induce the expression of early markers of bone differentiation (e.g. smad-1, smad-5, runx2/cbfa1, dlx5, osterix, osteopontin, bone sialoprotein and alkaline phosphatase) in C2C12 cells and in human adipose stem cells. The described approach is a promising method for producing large amounts of different recombinant BMPs that show potential for novel biomedical applications.