Osteogenesis coordinated in C3H10T1/2 cells by adipogenesis-dependent BMP-2 expression system.
A novel tissue engineering for bone formation has been proposed, to make osteoblast differentiation balanced by transfecting the mesenchymal stem cells with a gene encoding human bone morphogenetic protein-2 (hBMP-2) under the control of adipocyte specific lipoprotein lipase (LPL) promoter. Due to the promoter specificity, the initiation of BMP transcription is dependent on adipogenesis. For 14-day culture in the presence of ascorbic acid (asc) and beta-glycerophosphate (gly), nontransfected mouse embryonic fibroblast C3H10T1/2 (10T1/2) cells showed extensive accumulation of lipid droplets and adipocyte specific enzyme glycerol-3-phosphate dehydrogenase (G3PDH) mRNA expression, but exhibited neither BMP-2 expression, high alkaline phosphatase (ALP) activity which reflects osteoblast phenotype. On the other hand, transfected 10T1/2 cells showed hBMP-2 expression, high ALP activity and low level of G3PDH. mRNA expression accompanied with minimal lipid droplets. These results indicate that 10T1/2 cells are proved to be differentiated with maintaining coordinated balance of adipogenesis and osteogenesis, when they are transfected by the gene encoding hBMP-2 under the control of LPL promoter.
Mie
M
M
Department of Biological Information, Graduate School of Bioscience and Biotechnology, Tokyo Institute of Technology, Yokohama, Japan. maizawa@bio.titech.ac.jp
Ohgushi
H
H
Yanagida
Y
Y
Haruyama
T
T
Kobatake
E
E
Aizawa
M
M
D017667:Adipocytes / Q000166:cytology / Q000502:physiology*; D000818:Animals; D001205:Ascorbic Acid / Q000494:pharmacology; D001483:Base Sequence; D001698:Biomedical Engineering; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins / Q000235:genetics / Q000502:physiology*; D002454:Cell Differentiation; D002460:Cell Line; D017931:DNA Primers / Q000235:genetics; D015870:Gene Expression; D006801:Humans; D008071:Lipoprotein Lipase / Q000235:genetics; D051379:Mice; D010006:Osteoblasts / Q000166:cytology / Q000502:physiology; D010012:Osteogenesis / Q000502:physiology*; D011401:Promoter Regions, Genetic / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D014162:Transfection; D016212:Transforming Growth Factor beta
C525718:BMP2 protein, human; C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2; D019485:Bone Morphogenetic Proteins; D017931:DNA Primers; D012333:RNA, Messenger; D016212:Transforming Growth Factor beta; D008071:Lipoprotein Lipase; D001205:Ascorbic Acid