骨髓间充质干细胞向成牙本质样细胞分化的体外实验研究

In vitro study on the odontogenic differentiation potentiality of bone marrow mesenchymal stem cells

目的:观察骨髓间充质干细胞向成牙本质样细胞分化的能力。方法:分离培养SD大鼠骨髓间充质干细胞、胎鼠牙胚细胞,制备牙胚细胞条件培养液;用牙胚细胞条件培养液诱导SD大鼠骨髓间充质干细胞7d,通过免疫荧光染色和RT-PCR方法,观测其表达成牙本质细胞特异性标志物—牙本质涎蛋白(DSP)、牙本质基质蛋白1(DMP-1)的情况,明确骨髓间充质干细胞能否向成牙本质样细胞分化。结果:大鼠骨髓间充质干细胞在诱导体系中生长状态良好。培养7d后,部分细胞抗牙本质涎蛋白(DSP)、抗牙本质基质蛋白1(DMP-1)免疫荧光染色呈阳性;RT-PCR显示mRNA水平表达牙本质涎磷蛋白(DSPP)和牙本质基质蛋白(DMP-1)。结论:大鼠骨髓间充质干细胞在牙胚细胞条件培养液的诱导下可以分化为成牙本质样细胞。

AIM：To explore the possibility of bone marrow mesenchymal stem cells （BMSCs）differentiating into odontoblast-like cells.METHODS：BMSCs were isolated from two-week Sprague-Dawley rats and cultured in tooth germ cells conditioned medium （TGC-CM）.After 7 d induction,expression of dentin sialoprotein （DSP） and dentin matrix protein 1 （DMP1）,and specific markers of odontoblasts,were observed by immunofluorescence staining and RT-PCR.RESULTS：After 7 days induction in TGC-CM,positive staining of DSP and DMP-1 were detected in BMSCs by immunofluorescence staining.RT-PCR demonstrated mRNA expression of dentin sialophosphoprotein （DSPP） and DMP-1 by BMSCs.CONCLUSION：Rat BMSCs induced by TGC-CM could differentiate into odontoblast-like cells.