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Immortalization and characterization of mouse floxed Bmp2/4 osteoblasts |
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| Authors: | Li-An Wu Guohua Yuan Guobin Yang Iris Ortiz-Gonzalez Yong Cui Kevin J Donly Shuo Chen |
| Institution: | a Department of Pediatric Dentistry, The University of Texas Health Science Center at San Antonio, TX, USA b Department of Periodontics, Dental School, The University of Texas Health Science Center at San Antonio, TX, USA c Department of Pediatric Dentistry, School of Stomatology, The Fourth Military Medical University, Xi-an, China d Key Laboratory of Oral Biomedical Engineering Ministry of Education, Wuhan, China e Department of Oral/Maxillofacial Surgery, University of Alabama, Birmingham, AL, USA |
| Abstract: | 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. |
| Keywords: | Immortalization Floxed Bmp2/4 Gene expression Osteoblasts SV40 T-Ag |
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