title: Development and characterization of a mouse floxed Bmp2 osteoblast cell line that retains osteoblast genotype and phenotype.
abstract: Bone morphogenetic protein 2 (Bmp2) is essential for osteoblast differentiation and osteogenesis. Generation of floxed Bmp2 osteoblast cell lines is a valuable tool for studying the effects of Bmp2 on osteoblast differentiation and its signaling pathways during skeletal metabolism. Due to relatively limited sources of primary osteoblasts, we have developed cell lines that serve as good surrogate models for the study of osteoblast cell differentiation and bone mineralization. In this study, we established and characterized immortalized mouse floxed Bmp2 osteoblast cell lines. Primary mouse floxed Bmp2 osteoblasts were transfected with pSV3-neo and clonally selected. These transfected cells were verified by PCR and immunohistochemistry. To determine the genotype and phenotype of the immortalized cells, cell morphology, proliferation, differentiation and mineralization were analyzed. Also, expression of osteoblast-related gene markers including Runx2, Osx, ATF4, Dlx3, bone sialoprotein, dentin matrix protein 1, osteonectin, osteocalcin and osteopontin were examined by quantitative RT-PCR and immunohistochemistry. These results showed that immortalized floxed Bmp2 osteoblasts had a higher proliferation rate but preserved their genotypic and phenotypic characteristics similar to the primary cells. Thus, we, for the first time, describe the development of immortalized mouse floxed Bmp2 osteoblast cell lines and present a useful model to study osteoblast biology mediated by BMP2 and its downstream signaling transduction pathways.
authors.affiliation: Department of Pediatric Dentistry, Dental School, The University of Texas Health Science Center at San Antonio, 7703 Floyd Curl Drive, San Antonio, TX 78229-3900, USA.
authors.forename: Li-an
authors.initials: LA
authors.lastname: Wu
authors.forename: Junsheng
authors.initials: J
authors.lastname: Feng
authors.forename: Lynn
authors.initials: L
authors.lastname: Wang
authors.forename: Yan-dong
authors.initials: YD
authors.lastname: Mu
authors.forename: Andrew
authors.initials: A
authors.lastname: Baker
authors.forename: Kevin J
authors.initials: KJ
authors.lastname: Donly
authors.forename: Stephen E
authors.initials: SE
authors.lastname: Harris
authors.forename: Mary
authors.initials: M
authors.lastname: MacDougall
authors.forename: Shuo
authors.initials: S
authors.lastname: Chen
mesh_terms: D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics / Q000378:metabolism*; D002113:Calcification, Physiologic; D002454:Cell Differentiation; D002460:Cell Line; D049109:Cell Proliferation; D048430:Cell Shape; D005838:Genotype; D051379:Mice; D008822:Mice, Transgenic; D010006:Osteoblasts / Q000166:cytology* / Q000502:physiology*; D010641:Phenotype
chemical_list: C525719:Bmp2 protein, mouse; D055396:Bone Morphogenetic Protein 2