Immortalization and characterization of mouse floxed Bmp2/4 osteoblasts

Li An Wu, Guohua Yuan, Guobin Yang, Iris Ortiz-Gonzalez, Wuchen Yang, Yong Cui, Mary MacDougall, Kevin J. Donly, Stephen Harris, Shuo Chen

Research output: Contribution to journalArticlepeer-review

5 Scopus citations

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.

Original languageEnglish (US)
Pages (from-to)89-95
Number of pages7
JournalBiochemical and Biophysical Research Communications
Volume386
Issue number1
DOIs
StatePublished - Aug 14 2009

Keywords

  • Floxed Bmp2/4
  • Gene expression
  • Immortalization
  • Osteoblasts
  • SV40 T-Ag

ASJC Scopus subject areas

  • Biophysics
  • Biochemistry
  • Molecular Biology
  • Cell Biology

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