bFGF和地塞米松对骨髓基质干细胞增殖与分化的影响

Effects of Basic Fibroblast Growth Factor and Dexamethasone on Proliferation and Osteogenic Differentiation of Rat Bone Marrow-Derived Mesenchymal Stem Cells

目的:探讨碱性成纤维生长因子(bFGF)和地塞米松(Dex)在体外培养条件下对SD大鼠骨髓基质干细胞(MSC)增殖与分化的影响。方法:用DMEM冲洗股骨、胫骨骨髓腔,收集骨髓细胞悬液,接种在塑料培养瓶内进行体外培养及扩增,并分别加入不同浓度bFGF和Dex,通过MTT法检测2者对MSC增殖的影响,通过RT-PCR技术测定碱性磷酸酶(ALP)、骨钙素(OCN)和骨桥蛋白(OPN)的表达,了解2者对MSC分化的影响。结果:bFGF对MSC增殖呈剂量依赖性,10ng/ml时促增殖最明显;Dex对骨髓基质细胞的增殖起抑制作用,并随Dex浓度的升高而增强。bFGF降低MSC的ALP活性及OCN、OPN的表达;而Dex以及2者联合应用则增加ALP活性,促进MSC的ALP、OCN及OPN表达增加。结论:bFGF促进MSC增殖、抑制其分化;Dex抑制MSC增殖,但可促进其分化,bFGF(10ng/ml)和Dex(10-8mol/L)联合应用能有效的促进MSC的增殖和成骨分化。

Objective：To investigate in vitro effects of basic fibroblast growth factor （bFGF） and dexamethasone on the proliferation and differentiation of rat bone marrow derived mesenchymal stem cells （MSC）. Methods： MSC were isolated from young rat femur and tibia marrow flushed out with DMEM using a needle and syringe, and planted in plastic culture flask. The influence of different doses of bFGF and dexamethasone on proliferation of MSC was observed by MTT test. The expression of alkaline ghosphatase （ALP）, osteocalcin and osteopontin was tested by using RT PCR. Results： bFGF could promote the MSC proliferation in a dose-dependent manner, especially in 10 ng/ml, and dexamethasone could inhibit the proliferation of MSC in a concentration-dependent manner. BFGF decreased the expression of ALP, osteocalcin and osteopontin in MSC, while dexamethasone and combined use of them could increased the expression of ALP, osteocalcin and osteopontin in MSC. Conclusion： bFGF promoted the proliferation and inhibited the differentiation of MSC. Dexamethasone inhibited the proliferation of MSC, but induced them to differentiate into osteoblasts. Combined use of bFGF （10 ng/ml） with dexamethasone （10^-8 mol/L） can effectively stimulate MSC to proliferate and osteogenic differentiation.