title: FGF-2 abolishes the chondrogenic effect of combined BMP-6 and TGF-beta in human adipose derived stem cells.
abstract: In this study, we investigated the influence of transforming growth factor beta 3 (TGF-beta3), bone morphogenetic protein 6 (BMP-6) and basic fibroblast growth factor (FGF-2) on chondrogenesis in adipose derived stem cells (ASC). Cells were isolated from liposuction material, expanded and subjected to chondrogenic differentiation. Micromass pellets were cultured in chondrogenic medium containing 10 ng/mL TGF-beta3 which was additionally supplemented with 10 ng/mL BMP-6, 10 ng/mL FGF-2 or a combination of both. We quantitatively evaluated the cartilage specific gene expression after 14 days of culture. The end point measurements on day 35 included glycosaminoglycan (GAG) quantification, histological staining for chondrogenic markers, and transmission electron microscopy (TEM). In comparison to cultures induced with TGF-beta3/FGF-2, the presence of TGF-beta3/BMP-6 demonstrated strong induction of collagen type II, collagen type IX and aggrecan mRNA expression. This was corroborated by quantification and histological staining for GAGs and immunohistological staining for collagen II. However, when a combination of BMP-6 and FGF-2 in addition to TGF-beta3 was added, FGF-2 counteracted BMP-6, as indicated by reduced marker gene expression and weak to absent staining for GAGs. In conclusion, this study demonstrates that BMP-6 combined with TGF-beta3 is a potent inducer of chondrogenesis in human ASC. In contrast, FGF-2 does not contribute to differentiation, but rather suppresses the chondrogenic potential of BMP-6.
authors: Ludwig Boltzmann Institute for Experimental and Clinical Traumatology, AUVA Research Center, Linz/Vienna, Austria.
authors: Florian
authors: F
authors: Hildner
authors: Anja
authors: A
authors: Peterbauer
authors: Susanne
authors: S
authors: Wolbank
authors: Sylvia
authors: S
authors: Nürnberger
authors: Stefan
authors: S
authors: Marlovits
authors: Heinz
authors: H
authors: Redl
authors: Martijn
authors: M
authors: van Griensven
authors: Christian
authors: C
authors: Gabriel
mesh_terms: D000328:Adult; D000818:Animals; D015415:Biomarkers / Q000378:metabolism; D055418:Bone Morphogenetic Protein 6 / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects; D002478:Cells, Cultured; D020219:Chondrogenesis / Q000187:drug effects*; D003470:Culture Media / Q000737:chemistry; D005260:Female; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D006025:Glycosaminoglycans / Q000378:metabolism; D006801:Humans; D008875:Middle Aged; D012313:RNA / Q000378:metabolism; D013234:Stem Cells / Q000166:cytology / Q000187:drug effects* / Q000502:physiology*; D053782:Transforming Growth Factor beta3 / Q000494:pharmacology*
chemical_list: D015415:Biomarkers; D055418:Bone Morphogenetic Protein 6; D003470:Culture Media; D006025:Glycosaminoglycans; D053782:Transforming Growth Factor beta3; D016222:Fibroblast Growth Factor 2; D012313:RNA