人牙源性间充质细胞向成牙本质细胞的诱导分化

Differentiation induction of dental mesenchymal cells into odontoblasts

目的:建立人牙源性间充质细胞向成牙本质细胞分化的体外诱导方案。方法:应用bFGF+IGF 1或TGF β1分别对培养早期的人牙源性间充质细胞进行平面定向诱导,观察诱导后细胞的形态学改变,采用免疫荧光染色和RT PCR方法检测成牙本质细胞标志物———DSP蛋白和DSPPmRNA在诱导后细胞中的表达,并通过VonKossa染色检测诱导后细胞的矿化能力。结果:诱导后部分细胞出现单侧较长的细胞突起,表达DSP蛋白和DSPPmRNA,体外连续培养可自发形成矿化结节,出现较典型的成牙本质细胞的形态和功能特征。结论:bFGF+IGF 1或TGF β1可促进牙源性间充质细胞向成牙本质细胞分化。

Objective: To induce human dental mesenchymal cells to differentiate into odontoblasts in vitro.Methods:The cultured human dental mesenchymal cells were induced in two-dimensional culture model by bFGF(10 ng/ml)+IGF-1(100 ng/ml) or TGF-β1(5 ng/ml) for 4-7 d. Cell growth and morphology after induction were observed. The expression of human DSP protein and DSPP mRNA were detected by immunofluorescent staining and RT-PCR. Mineralization capability of the induced cells was evaluated using Von Kossa staining. Results:In both bFGF+IGF and TGF-β1 groups 20%-30% of the induced cells showed long single process.DSP protein and DSPP mRNA were observed in the induced cells.Mineralized nodules were found in the induction cultures.Conclusion: bFGF+IGF-1 or TGF-β1 can induce dental mesenchymal cells to differentiate into odontoblasts.