转化生长因子β3诱导人乳牙牙髓干细胞向成牙本质样细胞的分化（英文）  

Transforming growth factor beta 3 induced odontoblast-like differentiation of stem cells from human exfoliated deciduous teeth

作　　者：周慧 林锦梅[2] 任飞[3] 刘建平[4] 章锦才[3] 徐平平[3] 杨勤[3] 陈晓春[3] 

机构地区：[1]珠海市口腔医院,广东省珠海市519000 [2]广州医科大学,广东省广州市510182 [3]广东省口腔医院.南方医科大学附属口腔医院,广东省广州市510260 [4]中山大学孙逸仙纪念医院,广东省广州市510120

出　　处：《中国组织工程研究》2014年第23期3745-3750,共6页Chinese Journal of Tissue Engineering Research

基　　金：Major Diseases Prevention and Social Development Projects for Eugenics Technology of Guangdong Provincial Science and Technology Department,No.2009B030801253;the Zhuhai Health Bureau Medical Scientific Research Project,No.2013003~~

摘　　要：背景:转化生长因子β3与肝素联合作用后对人乳牙牙髓干细胞的增殖和矿化能力的作用目前仍有待研究。目的:探讨转化生长因子β3对人乳牙牙髓干细胞向成牙本质样细胞分化的作用。方法:采用酶消化法将人乳牙牙髓分离培养,获得人乳牙牙髓干细胞;对体外培养的第3代人乳牙牙髓干细胞单独加入25μg/L重组转化生长因子β3、10 U/mL肝素,或者二者联合进行培养;通过Q-PCR和Western-blotting方法,分别检测乳牙牙髓干细胞中牙本质涎磷蛋白基因及牙本质涎蛋白表达的情况;通过碱性磷酸酶试剂盒检测乳牙牙髓干细胞碱性磷酸酶活性的改变。结果与结论:乳牙牙髓干细胞在诱导体系中生长状态良好。25μg/L转化生长因子β3+10 U/mL肝素联合作用组的碱性磷酸酶活性明显增强,与转化生长因子β3单独作用组、肝素单独作用组以及对照组相比差异有显著性意义(P<0.01);转化生长因子β3+肝素联合作用组的茜素红染色呈强阳性,Q-PCR和Western-blotting结果均显示,转化生长因子β3+肝素联合作用组的牙本质涎磷蛋白mRNA和蛋白表达均明显升高。结果可见在转化生长因子β3与肝素联合作用的诱导下,可促进乳牙牙髓干细胞分化为成牙本质样细胞。BACKGROUND：Studies have reported that the superfamily of transforming growth factors exert a role in the mineralization of various stem cells, but the combination effects of transforming growth factorβ3 and heparin on proliferation and mineralization ability of stem cells from human exfoliated deciduous teeth remains to be studied. OBJECTIVE：To explore the effect of transforming growth factorβ3 on odontoblast-like differentiation of stem cells from human exfoliated deciduous teeth. METHODS：Human deciduous teeth were col ected using enzyme digestion. The 3rd cells were incubated with 25μg/L recombinant human transforming growth factorβ3, 10 U/mL heparin or their combination. The dentin sialophosphoprotein mRNA and dentinsialoprotein expressions were detected by Q-PCR and western blot assay. Alkaline phosphatase activity was determined using alkaline phosphatase kit. RESULTS AND CONCLUSION：Stem cells from human exfoliated deciduous teeth grew wel after induction. The activity of alkaline phosphatase in the combination group was significantly higher than that in the transforming growth factorβ3, heparin and control groups （P〈0.01）. After combination induction, the cells were strongly positive for alizarin red staining. Results fromα-PCR and western blot assay showed that the expressions of dentin sialophosphoprotein were both remarkably increased at mRNA and protein levels. In summary, stem cells from human exfoliated deciduous teeth can differentiate into odontoblast-like cells under the induction of transforming growth factorβ3 plus heparin.

关 键 词：干细胞 分化 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：R394.2[医药卫生—医学遗传学]