Induced differentiation of cultured rabbit mesenchymal stem cells by basic fibroblast growth factor and recombinant human bone morphogenetic protein-2

Yunsheng HU; Qingyu FAN; Baoan MA; Dianzhong ZHANG; Yunyan LIU

ABSTRACT

BACKGROUND:

Mesenchymal stem cells (MSCs) are capable of differentiating into a variety of lineages, including bone, cartilage, or fat, depending on the inducing stimuli, specific growth and differentiation factors.Recombinant human bone morphogenetic protein 2 (rhBMP-2) produced by gene engineering has an obvious osteoinductive activity and can induce undifferentiated mesenchymal cells into cartilage and bone irreversibly, resulting in new bone formation. Basic fibroblast growth factor (bFGF) modulates chondrogenic and osteogenic differentiation of MSCs.

OBJECTIVE:

To investigate the effect of bFGF and rhBMP-2 on the differentiation and proliferation of cultured rabbit mesenchymal stem cell in order to find out an optimal way of osteogenesis instead of conventional osteogenic supplements (OS).

DESIGN:

A randomized and controlled trial.

SETTING:

General Institute of Orthopaedic Oncology, Tangdu Hospital,Fourth Military Medical University of Chinese PLA.MATERIALS The subjects were rabbit mesenchymal stem cells cultured by the author.

METHODS:

This experiment was conducted at the Center of Orthopaedic Surgery, General Institute of Orthopaedic Oncology, Tangdu Hospital,Fourth Military Medical University of Chinese PLA from June 2004 to December 2004. ①Rabbit MSCs cultured in vitro were treated with different growth factor (100 μg/L rhBMP-2, 100 μg/L bFGF, 10 μg/L rhBMP-2and 100 μg/L bFGF and OS; ②The proliferation and differentiation of MSCs were observed through activity of MTT, expression of alkaline phosphatase(ALP), and von Kossa staining.MAIN OUTCOME

MEASURES:

the rate of proliferation and the activity of ALP.

RESULTS:

①rhBMP-2 could promote the proliferation and differentiation of MSCs, especially the cell differentiation; ②bFGF could stimulate the proliferation , the cellular proliferation rate increased 100% as compared with control group, and has no effect on differentiation of MSCs , but it could enhance effect on the cell proliferation of rhBMP-2.

CONCLUSION:

bFGF and rhBMP-2 are effective induction factors for MSCs. Both of them can stimulate the proliferation and differentiation of MSCs in vitro. bFGF and rhBMP-2 exerted a synergetic action in speeding up the pace of osteoinduction and osteogenesis and can be used to differentiate seed cells for tissue engineering bone.