MIP-3α 联合成骨诱导因子诱导大鼠脂肪干细胞向成牙本质样细胞分化*

目的:观察巨噬细胞炎性蛋白-3α(MIP-3α)对大鼠脂肪干细胞(Adipose derived stem cells,ASCs)向成牙本质样细胞体外分化作用的影响。方法:分离、培养并鉴定大鼠ASCs;以MIP-3α联合成骨诱导因子(地塞米松,β-甘油磷酸钠,以及抗坏血酸)诱导第3代大鼠ASCs向成牙本质样细胞定向分化。诱导培养1、4、7d后,分别测定碱性磷酸酶(alkaline phosphatese,ALP)活性,并用RT-PCR及Western Blot检测成牙本质细胞的标志基因dspp及标志物牙本质涎蛋白(DSP)。结果:与单独加入成骨诱导因子相比,MIP-3α与成骨诱导因子联合应用能使ALP活性、dspp的mRNA表达以及DSP升高。结论:本研究显示MIP-3α与成骨诱导因子联合应用可以增强成牙本质细胞相关基因以及蛋白的表达,为牙齿再生种子细胞的寻找开辟了一条新思路。

Induction of Odontoblast-Like Cells From Rat Adipose Derived Stem Cells by MIP-3 Combined with Osteogenic Factors in Vitro

Objective: To investigate the effect of MIP-3 on rat adipose derived stem cells’ differentiation into odontoblast-like cells in vitro.Method: Rat adipose-derived stem cells were isolated,cultured and their multi-potential differentiation capacities were analyzed.Osteogenic factors(dexamethasone,β-glycerophoshate and ascorbic acid) were used to induce rat ASCs to differentiated into odontoblast-like cells in the presence or absence of MIP-3.AT Day 1,4,and 7,ALP activity was measured,and dspp and DSP were examined by RT-PCR and Western Blotting,respectively.Results: The combination use of MIP-3 and osteogenic factors increased ALP activity,dspp expression,and DSP protein level as compared with osteogenic factos alone group.The control group showed the lowest ALP activity and no dspp expression or DSP protein synthesized.Conclusion: This study demonstrates that the combination use of MIP-3 and osteogenic factors may induce rat ASCs to differentiate towards odontoblast-like cells by promoting ALP activity and dspp expression and DSP protein level,and thus provide a new insight for the selection of tooth tissue engineering seed cells.