转化生长因子β3联合肝素在人乳牙牙髓干细胞向成牙本质样细胞分化中的作用  

Differentiation of odontoblast-like cells from human exfoliated deciduous teeth-derived stem cells by transforming growth factor β3 in combination with heparin in vitro

作　　者：任飞[1] 周慧 刘建平[3] 章锦才[1] 徐平平[1] 杨勤[1] 陈晓春[1] 

机构地区：[1]广东省口腔医院,南方医科大学附属口腔医院,广州市510260 [2]广东省珠海市口腔医院,519000 [3]中山大学孙逸仙纪念医院,广州市510120

出　　处：《实用医学杂志》2014年第12期1887-1890,共4页The Journal of Practical Medicine

摘　　要：目的:探讨转化生长因子β3(TGF-β3)诱导人乳牙牙髓干细胞(SHED)向成牙本质样细胞分化的能力。方法:采用酶消化法将人乳牙牙髓分离培养,获得SHED;对体外培养的第3代SHED分别单独加入25 ng/mL的重组TGF-β3,或与肝素联合进行培养;通过Q-PCR和Western-blotting方法,分别观察SHED表达成牙本质细胞特异性标志物-牙本质涎磷蛋白(DSPP)基因及其基因表达产物-牙本质涎蛋白(DSP)的情况;通过碱性磷酸酶(AKP)试剂盒检测SHED的AKP活性的改变;细胞爬片行免疫化学染色和茜素红染色。结果:SHED在诱导体系中生长状态良好。25 ng/mL TGF-β3联合10 U/mL肝素作用组的AKP活性明显增强,与TGF-β3单独作用组、肝素单独作用组以及对照组相比差异具有统计学意义(P<0.01);TGF-β3联合肝素作用组的茜素红染色呈强阳性,Q-PCR和Western-blotting结果均显示,该组的DSPP表达在mRNA水平和蛋白质水平上均明显升高。结论:在TGF-β3与肝素联合作用的诱导下,可促进SHED分化为成牙本质样细胞。Objective To investigate the ability of human exfoliated deciduous teeth-derived stem cells （SHED） to differentiate into odontoblast-like cells. Methods SHEDs were isolated by enzyme digestion method. The 3nd passage SHEDs were incubated with 25 ng/mL recombinant human TGF-β3, or with TGF-β3 in combination with heparin. The DSPP expression was detected by Q-PCR and Western-blotting assay. Alizarin red staining, immunhistochemistry assay and alkaline phosphatase （AKP） activity assay were performed, respectively. Result The AKP activity was enhance by TGF-β3 in combination with heparin. Alizarin red staining was positive in TGF-β3-heparin groups, with the increase of DSPP expression at both mRNA and protein level. Conclusion TGF-β3 in combination with heparin can enhance the differentiation of human exfoliated deciduous teeth-derived stem cells into odontoblast-like cells.

关 键 词：人乳牙牙髓干细胞 转化生长因子Β3 肝素 成牙本质样细胞 

分 类 号：R780.2[医药卫生—口腔医学]