title: [Bone marrow stromal cells co-modified with BMP-2 and bFGF gene].
abstract: OBJECTIVE
  To establish modified bone marrow stromal cells(BMSCs) which can express BMP-2 and bFGF stably.
  
  
  METHODS
  BMP-2 and bFGF gene were amplified by RT-PCR, and then cloned into the expression vector pcDNA3.0. After being confirmed by DNA sequencing, pcDNA3.0-BMP-2 and pcDNA3.0-bFGF were co-transfected into rat BMSCs with Lipofectamine 2000 reagent. The expression of BMP-2 and bFGF gene in rat BMSCs was detected by RT-PCR, Western blot, immunohistochemical staining and ELISA.
  
  
  RESULTS
  BMP-2 and bFGF gene were cloned, and their sequences were identical with those in GenBank. The expression plasmids, pcDNA3.0-BMP-2 and pcDNA3.0-bFGF, were constructed and co-transfected into rMSCs successfully. RT-PCR showed the mass transcription of BMP-2 and bFGF mRNA in transfected BMSCs. Western blot, immunohistochemical staining and ELISA confirmed the expression of BMP-2 and bFGF genes in transfected cells and in the supernatant.
  
  
  CONCLUSIONS
  We have constructed the optimal rat BMSCs which can be used in bone tissue engineering.
authors.affiliation: Institute of Biomedical Engineering, West China Medical Center of Sichuan University, Chengdu 610041, China. yi_lee0103@yahoo.com.cn
authors.forename: Yi
authors.initials: Y
authors.lastname: Li
authors.forename: Huai-qing
authors.initials: HQ
authors.lastname: Chen
authors.forename: Min
authors.initials: M
authors.lastname: Cheng
authors.forename: Yan-juan
authors.initials: YJ
authors.lastname: Tang
mesh_terms: D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology*; D055396:Bone Morphogenetic Protein 2 / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D016222:Fibroblast Growth Factor 2 / Q000235:genetics / Q000378:metabolism*; D015870:Gene Expression; D018014:Gene Transfer Techniques; D051381:Rats; D017207:Rats, Sprague-Dawley; D017154:Stromal Cells / Q000378:metabolism*
chemical_list: D055396:Bone Morphogenetic Protein 2; D016222:Fibroblast Growth Factor 2