[Effects of BMP-2 and FGF-2 on osteoblast differentiation of murine MSCs in vitro].
OBJECTIVE
To study the effects of BMP-2 and FGF-2 on osteoblast differentiation of murine MSCs in vitro.


METHODS
The bone marrow cells were collected from 3-18 month old C57BL/6J mice (50 mice), and they were isolated, enriched and expanded using bone marrow adherent culture, and then purified by immunomagnetic microbeads. At last they were identified as mesenchymal stem cells (MSCs). After the MSCs were cultured adherently for 24 hours, 100 microg/L BMP-2 and 0.5 nmol/L FGF-2 were added into osteogenic media separately for 7, 14 and 21 days, alkaline phosphatase (ALP) staining, alkaline phosphatase activities, Vonkossa staining, and Alizarin red S staining were performed step by step. Osteoblast differentiation marker genes including Runx2/cbfa1, ALP, collagen-1, and osteocalcin were investigated by RT real time PCR.


RESULTS
ALP activities, calcium deposition and the osteoblast differentiation marker genes of BMP-2 group were markedly higher than those of osteogenic group. The mRNA of Runx2/cbfa1, ALP, collagen-1, and osteocalcin of BMP-2 group was highly expressed than that of osteogenic group. It was the same with FGF-2 group, but less evident than BMP-2 group.


CONCLUSIONS
BMP-2 and FGF-2 can induce murine MSCs to differentiate into osteoblast at different degrees.
Zhang
Wei-xi
WX
Department of Neurology, First Hospital of Sun Yat-sen University, Guangzhou 510080, China. weixizhang@hotmail.com
Chen
Song-lin
SL
Yao
Xiao-li
XL
Lu
Xi-lin
XL
Zhou
Chen
C
Leng
Yan
Y
Shi
Xingming
X
D048868:Adaptor Proteins, Signal Transducing / Q000235:genetics; D000818:Animals; D001854:Bone Marrow Cells / Q000166:cytology; D055396:Bone Morphogenetic Protein 2 / Q000494:pharmacology*; D002454:Cell Differentiation / Q000187:drug effects*; D003094:Collagen / Q000235:genetics; D050659:Core Binding Factor Alpha 1 Subunit / Q000235:genetics; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D008565:Membrane Proteins; D059630:Mesenchymal Stem Cells / Q000166:cytology* / Q000187:drug effects* / Q000378:metabolism; D051379:Mice; D008810:Mice, Inbred C57BL; D010006:Osteoblasts / Q000166:cytology* / Q000187:drug effects / Q000378:metabolism; D015675:Osteocalcin / Q000235:genetics; D010012:Osteogenesis / Q000187:drug effects*; D016133:Polymerase Chain Reaction
D048868:Adaptor Proteins, Signal Transducing; C480020:Atrnl1 protein, mouse; D055396:Bone Morphogenetic Protein 2; D050659:Core Binding Factor Alpha 1 Subunit; D008565:Membrane Proteins; D016222:Fibroblast Growth Factor 2; D015675:Osteocalcin; D003094:Collagen