Co-stimulation with bone morphogenetic protein-9 and FK506 induces remarkable osteoblastic differentiation in rat dedifferentiated fat cells.
Dedifferentiated fat (DFAT) cells, which are isolated from mature adipocytes using the ceiling culture method, exhibit similar characteristics to mesenchymal stem cells, and possess adipogenic, osteogenic, chondrogenic, and myogenic potentials. Bone morphogenetic protein (BMP)-2 and -9, members of the transforming growth factor-β superfamily, exhibit the most potent osteogenic activity of this growth factor family. However, the effects of BMP-2 and BMP-9 on the osteogenic differentiation of DFAT remain unknown. Here, we examined the effects of BMP-2 and BMP-9 on osteoblastic differentiation of rat DFAT (rDFAT) cells in the presence or absence of FK506, an immunosuppressive agent. Co-stimulation with BMP-9 and FK506 induced gene expression of runx2, osterix, and bone sialoprotein, and ALP activity compared with BMP-9 alone, BMP-2 alone and BMP-2+FK506 in rDFAT cells. Furthermore, it caused mineralization of cultures and phosphorylation of smad1/5/8, compared with BMP-9 alone. The ALP activity induced by BMP-9+FK506 was not influenced by addition of noggin, a BMP antagonist. Our data suggest that the combination of BMP-9 and FK506 potently induces osteoblastic differentiation of rDFAT cells.
Department of Periodontology, Kagoshima University, Graduate School of Medical and Dental Sciences, Kagoshima, Japan.
Toshiaki
T
Nakamura
Yukiya
Y
Shinohara
Sawako
S
Momozaki
Takehiko
T
Yoshimoto
Kazuyuki
K
Noguchi
D017667:Adipocytes / Q000502:physiology; D000469:Alkaline Phosphatase; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000494:pharmacology; D002352:Carrier Proteins / Q000494:pharmacology; D054337:Cell Dedifferentiation; D002454:Cell Differentiation / Q000187:drug effects* / Q000502:physiology; D055427:Growth Differentiation Factor 2 / Q000494:pharmacology*; D006801:Humans; D008297:Male; D010012:Osteogenesis / Q000187:drug effects*; D012333:RNA, Messenger / Q000378:metabolism; D051381:Rats; D017208:Rats, Wistar; D016559:Tacrolimus / Q000494:pharmacology*; D000099176:Noggin Protein
D000469:Alkaline Phosphatase; D055396:Bone Morphogenetic Protein 2; D002352:Carrier Proteins; D055427:Growth Differentiation Factor 2; D012333:RNA, Messenger; D016559:Tacrolimus; D000099176:Noggin Protein; C525718:BMP2 protein, human
BMP-9; Dedifferentiated fat cells; FK506; Osteoblastic differentiation; Smad