Cargando…

A Novel Human TGF-β1 Fusion Protein in Combination with rhBMP-2 Increases Chondro-Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells

Transforming growth factor-beta (TGF-β) is involved in processes related to the differentiation and maturation of osteoprogenitor cells into osteoblasts. Rat bone marrow (BM) cells were cultured in a collagen-gel containing 0.5% fetal bovine serum (FBS) for 10 days in the presence of rhTGF (recombin...

Descripción completa

Detalles Bibliográficos
Autores principales: Claros, Silvia, Rico-Llanos, Gustavo A., Becerra, José, Andrades, José A.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2014
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4139781/
https://www.ncbi.nlm.nih.gov/pubmed/24968268
http://dx.doi.org/10.3390/ijms150711255
_version_ 1782331409541627904
author Claros, Silvia
Rico-Llanos, Gustavo A.
Becerra, José
Andrades, José A.
author_facet Claros, Silvia
Rico-Llanos, Gustavo A.
Becerra, José
Andrades, José A.
author_sort Claros, Silvia
collection PubMed
description Transforming growth factor-beta (TGF-β) is involved in processes related to the differentiation and maturation of osteoprogenitor cells into osteoblasts. Rat bone marrow (BM) cells were cultured in a collagen-gel containing 0.5% fetal bovine serum (FBS) for 10 days in the presence of rhTGF (recombinant human TGF)-β1-F2, a fusion protein engineered to include a high-affinity collagen-binding decapeptide derived from von Willebrand factor. Subsequently, cells were moderately expanded in medium with 10% FBS for 4 days and treated with a short pulse of rhBMP (recombinant human bone morphogenetic protein)-2 for 4 h. During the last 2 days, dexamethasone and β-glycerophosphate were added to potentiate osteoinduction. Concomitant with an up-regulation of cell proliferation, DNA synthesis levels were determined. Polymerase chain reaction was performed to reveal the possible stemness of these cells. Osteogenic differentiation was evaluated in terms of alkaline phosphatase activity and mineralized matrix formation as well as by mRNA expression of osteogenic marker genes. Moreover, cells were placed inside diffusion chambers and implanted subcutaneously into the backs of adult rats for 4 weeks. Histological study provided evidence of cartilage and bone-like tissue formation. This experimental procedure is capable of selecting cell populations from BM that, in the presence of rhTGF-β1-F2 and rhBMP-2, achieve skeletogenic potential in vitro and in vivo.
format Online
Article
Text
id pubmed-4139781
institution National Center for Biotechnology Information
language English
publishDate 2014
publisher MDPI
record_format MEDLINE/PubMed
spelling pubmed-41397812014-08-21 A Novel Human TGF-β1 Fusion Protein in Combination with rhBMP-2 Increases Chondro-Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells Claros, Silvia Rico-Llanos, Gustavo A. Becerra, José Andrades, José A. Int J Mol Sci Article Transforming growth factor-beta (TGF-β) is involved in processes related to the differentiation and maturation of osteoprogenitor cells into osteoblasts. Rat bone marrow (BM) cells were cultured in a collagen-gel containing 0.5% fetal bovine serum (FBS) for 10 days in the presence of rhTGF (recombinant human TGF)-β1-F2, a fusion protein engineered to include a high-affinity collagen-binding decapeptide derived from von Willebrand factor. Subsequently, cells were moderately expanded in medium with 10% FBS for 4 days and treated with a short pulse of rhBMP (recombinant human bone morphogenetic protein)-2 for 4 h. During the last 2 days, dexamethasone and β-glycerophosphate were added to potentiate osteoinduction. Concomitant with an up-regulation of cell proliferation, DNA synthesis levels were determined. Polymerase chain reaction was performed to reveal the possible stemness of these cells. Osteogenic differentiation was evaluated in terms of alkaline phosphatase activity and mineralized matrix formation as well as by mRNA expression of osteogenic marker genes. Moreover, cells were placed inside diffusion chambers and implanted subcutaneously into the backs of adult rats for 4 weeks. Histological study provided evidence of cartilage and bone-like tissue formation. This experimental procedure is capable of selecting cell populations from BM that, in the presence of rhTGF-β1-F2 and rhBMP-2, achieve skeletogenic potential in vitro and in vivo. MDPI 2014-06-25 /pmc/articles/PMC4139781/ /pubmed/24968268 http://dx.doi.org/10.3390/ijms150711255 Text en © 2014 by the authors; licensee MDPI, Basel, Switzerland. http://creativecommons.org/licenses/by/3.0/ This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution license (http://creativecommons.org/licenses/by/3.0/).
spellingShingle Article
Claros, Silvia
Rico-Llanos, Gustavo A.
Becerra, José
Andrades, José A.
A Novel Human TGF-β1 Fusion Protein in Combination with rhBMP-2 Increases Chondro-Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells
title A Novel Human TGF-β1 Fusion Protein in Combination with rhBMP-2 Increases Chondro-Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells
title_full A Novel Human TGF-β1 Fusion Protein in Combination with rhBMP-2 Increases Chondro-Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells
title_fullStr A Novel Human TGF-β1 Fusion Protein in Combination with rhBMP-2 Increases Chondro-Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells
title_full_unstemmed A Novel Human TGF-β1 Fusion Protein in Combination with rhBMP-2 Increases Chondro-Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells
title_short A Novel Human TGF-β1 Fusion Protein in Combination with rhBMP-2 Increases Chondro-Osteogenic Differentiation of Bone Marrow Mesenchymal Stem Cells
title_sort novel human tgf-β1 fusion protein in combination with rhbmp-2 increases chondro-osteogenic differentiation of bone marrow mesenchymal stem cells
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC4139781/
https://www.ncbi.nlm.nih.gov/pubmed/24968268
http://dx.doi.org/10.3390/ijms150711255
work_keys_str_mv AT clarossilvia anovelhumantgfb1fusionproteinincombinationwithrhbmp2increaseschondroosteogenicdifferentiationofbonemarrowmesenchymalstemcells
AT ricollanosgustavoa anovelhumantgfb1fusionproteinincombinationwithrhbmp2increaseschondroosteogenicdifferentiationofbonemarrowmesenchymalstemcells
AT becerrajose anovelhumantgfb1fusionproteinincombinationwithrhbmp2increaseschondroosteogenicdifferentiationofbonemarrowmesenchymalstemcells
AT andradesjosea anovelhumantgfb1fusionproteinincombinationwithrhbmp2increaseschondroosteogenicdifferentiationofbonemarrowmesenchymalstemcells
AT clarossilvia novelhumantgfb1fusionproteinincombinationwithrhbmp2increaseschondroosteogenicdifferentiationofbonemarrowmesenchymalstemcells
AT ricollanosgustavoa novelhumantgfb1fusionproteinincombinationwithrhbmp2increaseschondroosteogenicdifferentiationofbonemarrowmesenchymalstemcells
AT becerrajose novelhumantgfb1fusionproteinincombinationwithrhbmp2increaseschondroosteogenicdifferentiationofbonemarrowmesenchymalstemcells
AT andradesjosea novelhumantgfb1fusionproteinincombinationwithrhbmp2increaseschondroosteogenicdifferentiationofbonemarrowmesenchymalstemcells