[Bone marrow stromal cells co-modified with BMP-2 and bFGF gene].
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.
Li
Yi
Y
Institute of Biomedical Engineering, West China Medical Center of Sichuan University, Chengdu 610041, China. yi_lee0103@yahoo.com.cn
Chen
Huai-qing
HQ
Cheng
Min
M
Tang
Yan-juan
YJ
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*
D055396:Bone Morphogenetic Protein 2; D016222:Fibroblast Growth Factor 2