Effects of BMP-2 and FGF-2 on osteoblast differentiation of murine MSCs in vitro

Wei xi Zhang, Song lin Chen, Xiao li Yao, Xi lin Lu, Chen Zhou, Yan Leng, Xing Ming Shi

Research output: Contribution to journalArticle

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Abstract

AIM: 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. CONCLUSION: BMP-2 and FGF-2 can induce murine MSCs to differentiate into osteoblast at different degrees.

Original languageEnglish (US)
Pages (from-to)1062-1065
Number of pages4
JournalXi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology
Volume24
Issue number11
StatePublished - Jan 1 2008
Externally publishedYes

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Fibroblast Growth Factor 2
Osteoblasts
Mesenchymal Stromal Cells
Alkaline Phosphatase
Osteocalcin
Differentiation Antigens
Staining and Labeling
Collagen
Microspheres
Inbred C57BL Mouse
Bone Marrow Cells
Genes
Real-Time Polymerase Chain Reaction
Bone Marrow
In Vitro Techniques
Calcium
Messenger RNA

ASJC Scopus subject areas

  • Medicine(all)

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Effects of BMP-2 and FGF-2 on osteoblast differentiation of murine MSCs in vitro. / Zhang, Wei xi; Chen, Song lin; Yao, Xiao li; Lu, Xi lin; Zhou, Chen; Leng, Yan; Shi, Xing Ming.

In: Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology, Vol. 24, No. 11, 01.01.2008, p. 1062-1065.

Research output: Contribution to journalArticle

Zhang, Wei xi ; Chen, Song lin ; Yao, Xiao li ; Lu, Xi lin ; Zhou, Chen ; Leng, Yan ; Shi, Xing Ming. / Effects of BMP-2 and FGF-2 on osteoblast differentiation of murine MSCs in vitro. In: Xi bao yu fen zi mian yi xue za zhi = Chinese journal of cellular and molecular immunology. 2008 ; Vol. 24, No. 11. pp. 1062-1065.
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AU - Leng, Yan

AU - Shi, Xing Ming

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N2 - AIM: 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. CONCLUSION: BMP-2 and FGF-2 can induce murine MSCs to differentiate into osteoblast at different degrees.

AB - AIM: 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. CONCLUSION: BMP-2 and FGF-2 can induce murine MSCs to differentiate into osteoblast at different degrees.

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