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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。
Objective: To investigate the effect of MIP-3α on the differentiation of rat adipose derived stem cells (ADSCs) into odontoblast-like cells in vitro. METHODS: Rat ASCs were isolated, cultured and identified. The third generation rat ASCs were induced to differentiate into odontoblast-like cells by MIP-3α combined with osteogenic factors (dexamethasone, β-glycerophosphate and ascorbic acid). The alkaline phosphatase (ALP) activity was measured after induction for 1, 4 and 7 days. The marker dsDNA and marker dentin sialoprotein (DSP) of odontoblast were detected by RT-PCR and Western Blot respectively. RESULTS: The combination of MIP-3α and osteoinductive factor increased ALP activity, dspp mRNA expression, and DSP elevation compared with osteogenic induction alone. Conclusion: This study shows that the combination of MIP-3α and osteoinductive factor can enhance the expression of odontoblast-related genes and proteins, which opens up a new idea for searching for regenerated seed cells.