TY - JOUR
T1 - Immortalization and characterization of mouse floxed Bmp2/4 osteoblasts
AU - Wu, Li An
AU - Yuan, Guohua
AU - Yang, Guobin
AU - Ortiz-Gonzalez, Iris
AU - Yang, Wuchen
AU - Cui, Yong
AU - MacDougall, Mary
AU - Donly, Kevin J.
AU - Harris, Stephen
AU - Chen, Shuo
N1 - Funding Information:
This work was supported in part by National Institutes of Health Grants DE019802 (S.C.), AR054616 (S.H.) and the grant from the Natural Science Foundation of China 30801293 (LA Wu) and 2009-10 3M ESPE Preventative Pediatric Dentistry Postdoctoral Research Fellowship of AAPD (LA Wu).
PY - 2009/8/14
Y1 - 2009/8/14
N2 - 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.
AB - 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.
KW - Floxed Bmp2/4
KW - Gene expression
KW - Immortalization
KW - Osteoblasts
KW - SV40 T-Ag
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U2 - 10.1016/j.bbrc.2009.05.144
DO - 10.1016/j.bbrc.2009.05.144
M3 - Article
C2 - 19501574
AN - SCOPUS:67649213889
SN - 0006-291X
VL - 386
SP - 89
EP - 95
JO - Biochemical and Biophysical Research Communications
JF - Biochemical and Biophysical Research Communications
IS - 1
ER -