title: BMP-4 increases activin A gene expression during osteogenic differentiation of mouse embryonic stem cells.
abstract: Activin A is a growth factor released by mature osteoblasts that has a critical effect on bone formation. We investigated the effect of bone morphogenetic protein (BMP)-4 on activin A gene expression during in vitro osteogenic differentiation of mouse embryonic stem (ES) cells. Embryoid bodies were cultured in retinoic acid (RA) for three days and then without RA for two days. Seeded cells received osteogenic medium with β-glycerophosphate, L-ascorbic acid 2-phosphate and dexamethasone during 19 days, with or without BMP-4. Six independent experiments were carried out. Real-time PCR was used to detect gene expression of activin A, Oct-4, Nanog, osteocalcin, RUNX2 and bone alkaline phosphatase. Immunofluorescence was used to co-localize activin A with the undifferentiation marker stage-specific embryonic antigen 1. Cells treated with BMP-4 had an increased gene expression of activin A, osteocalcin and bone alkaline phosphatase (p < 0.05). In conclusion, BMP-4 increases activin A gene expression during mouse ES cell differentiation into bone precursors.
authors.affiliation: Department of Obstetrics and Gynecology and.
authors.forename: Bruno M
authors.initials: BM
authors.lastname: Camargos
authors.forename: Rubens L C
authors.initials: RL
authors.lastname: Tavares
authors.forename: Helen L
authors.initials: HL
authors.lastname: Del Puerto
authors.forename: Luciana O
authors.initials: LO
authors.lastname: Andrade
authors.forename: Aroldo F
authors.initials: AF
authors.lastname: Camargos
authors.forename: Fernando M
authors.initials: FM
authors.lastname: Reis
mesh_terms: D028341:Activins / Q000378:metabolism*; D000818:Animals; D001205:Ascorbic Acid / Q000008:administration & dosage / Q000031:analogs & derivatives; D055415:Bone Morphogenetic Protein 4 / Q000494:pharmacology*; D002454:Cell Differentiation; D003470:Culture Media; D017931:DNA Primers / Q000235:genetics; D003907:Dexamethasone / Q000737:chemistry; D005347:Fibroblasts / Q000378:metabolism; D018507:Gene Expression Regulation, Developmental; D005994:Glycerophosphates / Q000737:chemistry; D051379:Mice; D008856:Microscopy, Fluorescence; D000066450:Mouse Embryonic Stem Cells / Q000166:cytology*; D010012:Osteogenesis / Q000187:drug effects*; D012333:RNA, Messenger / Q000378:metabolism; D060888:Real-Time Polymerase Chain Reaction; D014212:Tretinoin / Q000737:chemistry
chemical_list: C525810:Bmp4 protein, mouse; D055415:Bone Morphogenetic Protein 4; D003470:Culture Media; D017931:DNA Primers; D005994:Glycerophosphates; D012333:RNA, Messenger; C422405:activin A; D028341:Activins; C011669:ascorbate-2-phosphate; D014212:Tretinoin; D003907:Dexamethasone; D001205:Ascorbic Acid; C031463:beta-glycerophosphoric acid
keywords: BMP-4; differentiation; embryonic stem cells; osteogenesis; osteogenic precursors