Immortalization and characterization of mouse floxed Bmp2/4 osteoblasts.
Generation of a floxed Bmp2/4 osteoblast cell line is a valuable tool for studying the modulatory effects of Bmp2 and Bmp4 on osteoblast differentiation as well as relevant molecular events. In this study, primary floxed Bmp2/4 mouse osteoblasts were cultured and transfected with simian virus 40 large T-antigen. Transfection was verified by polymerase chain reaction (PCR) and immunohistochemistry. To examine the characteristics of the transfected cells, morphology, proliferation and mineralization were analyzed, expression of cell-specific genes including Runx2, ATF4, Dlx3, Osx, dentin matrix protein 1, bone sialoprotein, osteopontin, osteocalcin, osteonectin and collagen type I was detected. These results show that transfected floxed Bmp2/4 osteoblasts bypassed senescence with a higher proliferation rate, but retain the genotypic and phenotypic characteristics similar to the primary cells. Thus, we for the first time demonstrate the establishment of an immortalized mouse floxed Bmp2/4 osteoblast cell line.
Wu
Li-An
LA
Department of Pediatric Dentistry, The University of Texas Health Science Center at San Antonio, TX 78229-3900, USA.
Yuan
Guohua
G
Yang
Guobin
G
Ortiz-Gonzalez
Iris
I
Yang
Wuchen
W
Cui
Yong
Y
MacDougall
Mary
M
Donly
Kevin J
KJ
Harris
Stephen
S
Chen
Shuo
S
D000469:Alkaline Phosphatase / Q000096:biosynthesis; D000818:Animals; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics / Q000378:metabolism*; D055415:Bone Morphogenetic Protein 4 / Q000235:genetics / Q000378:metabolism*; D002113:Calcification, Physiologic; D002460:Cell Line; D002461:Cell Line, Transformed; D049109:Cell Proliferation; D005786:Gene Expression Regulation; D051379:Mice; D008817:Mice, Mutant Strains; D010006:Osteoblasts / Q000166:cytology / Q000378:metabolism*; D014162:Transfection
C525719:Bmp2 protein, mouse; C525810:Bmp4 protein, mouse; D055396:Bone Morphogenetic Protein 2; D055415:Bone Morphogenetic Protein 4; D000469:Alkaline Phosphatase