title: BMP7 promotes adipogenic but not osteo-/chondrogenic differentiation of adult human bone marrow-derived stem cells in high-density micro-mass culture.
abstract: The objective of our study was to elucidate the potential of bone morphogenetic protein-7 (BMP7) to initiate distinct mesenchymal lineage development of human adult mesenchymal stem cells (MSC) in three-dimensional micro-mass culture. Expanded MSC were cultured in high-density micro-masses under serum-free conditions that favor chondrogenic differentiation and were stimulated with 50-200 ng/ml BMP7 or 10 ng/ml transforming growth factor-beta3 (TGFbeta3) as control. Histological staining of proteoglycan with alcian blue, mineralized matrix according to von Kossa, and lipids with Oil Red O, immunostaining of type II collagen as well as real-time gene expression analysis of typical chondrogenic, adipogenic, and osteogenic marker genes showed that BMP7 promoted adipogenic differentiation of MSC. Micro-masses stimulated with BMP7 developed adipocytic cells filled with lipid droplets and showed an enhanced expression of the adipocyte marker genes fatty acid binding protein 4 (FABP4) and the adipose most abundant transcript 1 (apM1). Development along the chondrogenic lineage or stimulation of osteogenic differentiation were not evident upon stimulation with BMP7 in different concentrations. In contrast, TGFbeta3 directed MSC to form a cartilaginous matrix that is rich in proteoglycan and type II collagen. Gene expression analysis of typical chondrocyte marker genes like cartilage oligomeric matrix protein (COMP), link protein, aggrecan, and types IIalpha1 and IXalpha3 collagen confirmed chondrogenic differentiation of MSC treated with TGFbeta3. These results suggest that BMP7 promotes the adipogenic and not the osteogenic or chondrogenic lineage development of human stem cells when assembled three-dimensionally in micro-masses.
authors.affiliation: TransTissue Technologies GmbH, Tucholskystrasse 2, 10117 Berlin, Germany.
authors.forename: Katja
authors.initials: K
authors.lastname: Neumann
authors.forename: Michaela
authors.initials: M
authors.lastname: Endres
authors.forename: Jochen
authors.initials: J
authors.lastname: Ringe
authors.forename: Bernd
authors.initials: B
authors.lastname: Flath
authors.forename: Rudi
authors.initials: R
authors.lastname: Manz
authors.forename: Thomas
authors.initials: T
authors.lastname: Häupl
authors.forename: Michael
authors.initials: M
authors.lastname: Sittinger
authors.forename: Christian
authors.initials: C
authors.lastname: Kaps
mesh_terms: D050156:Adipogenesis; D000273:Adipose Tissue / Q000378:metabolism*; D000368:Aged; D001854:Bone Marrow Cells / Q000166:cytology*; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins / Q000378:metabolism / Q000502:physiology*; D002454:Cell Differentiation; D002478:Cells, Cultured; D019902:Chondrocytes / Q000378:metabolism*; D050556:Fatty Acid-Binding Proteins / Q000378:metabolism; D020869:Gene Expression Profiling; D006801:Humans; D008875:Middle Aged; D010006:Osteoblasts / Q000378:metabolism*; D013234:Stem Cells / Q000166:cytology*; D016212:Transforming Growth Factor beta / Q000378:metabolism / Q000502:physiology*; D053782:Transforming Growth Factor beta3 / Q000378:metabolism
chemical_list: C525819:BMP7 protein, human; D055419:Bone Morphogenetic Protein 7; D019485:Bone Morphogenetic Proteins; C493410:FABP4 protein, human; D050556:Fatty Acid-Binding Proteins; D016212:Transforming Growth Factor beta; D053782:Transforming Growth Factor beta3