rhBMP2对人牙髓干细胞分化及Delta蛋白表达的影响

doi:重庆市科技厅自然科学基金资助课题（CQCSTC

rhBMP₂对人牙髓干细胞分化及Delta蛋白表达的影响

于娜,何飞，谭颖徽

(第三军医大学新桥医院口腔科,重庆400037)

收稿日期:2008-12-11 修回日期:1900-01-01 出版日期:2009-03-28 发布日期:2009-08-14
通讯作者: 谭颖徽

Effects of rhBMP₂ on differentiation of human dental pulp stem cells and expression of Delta protein in vitro

YU Na, HE Fei, TAN Ying-hui

(Department of Stomatology, Xinqiao Hospital,Third Military Medical University, Chongqing 400037, China)

Received:2008-12-11 Revised:1900-01-01 Online:2009-03-28 Published:2009-08-14
Contact: TAN Ying-hui

摘要/Abstract

摘要： 目的：研究重组人骨形成蛋白2（rhBMP₂）对人牙髓干细胞分化及Delta蛋白表达的影响，为人牙髓干细胞的研究提供理论依据。方法：酶消化法获得人牙髓干细胞,光镜下进行形态学观察,免疫荧光法检测细胞表面标志的表达；取第5代牙髓干细胞, 分为实验组（加含50 μg•L^-1rhBMP₂的培养液）及阴性对照组（只加入培养液），检测碱性磷酸酶活性和Delta蛋白表达的变化。结果：人牙髓干细胞呈集落状生长, 免疫组织化学检测显示nestin、 vimentin染色呈阳性，免疫荧光法检测STRO-1呈阳性。与阴性对照组比较，实验组第7、14及21天碱性磷酸酶活性均明显升高（P<0.01）,第21天Western blotting法检测Delta蛋白表达明显升高(P<0.05)。结论：培养的牙髓干细胞具有干细胞特性，并且rhBMP₂可使其碱性磷酸酶活性增高并上调Delta蛋白表达，提示Delta蛋白可能参与牙髓干细胞向成牙本质样细胞分化的过程。

关键词: Delta蛋白, 牙髓干细胞

Abstract: Abstract:Objective To investigate the effects of rhBMP₂ on the differentiation of human dental pulp stem cells and expression of Delta protein in vitro. provide a theoretical basis for research on human dental pulp stem cells. Methods Monocell suspension was separated from the human adult dental pulp with collagenase Ⅰ and dispase digestion.Clonogenic cells were observed under light microscope and the expressions of surface markers were determined with immunofluorescence. divided into experimental group (containing 50 μg•L^-1rhBMP₂ in the culture medium) and negative control group (containing culture medium only).alkaline phosphatase and the expression of Delta proterin of the cells at at passage 5 were detected.Results The human dental pulp stem cells showed colony growth,the nestin and vimentin staining were positive by immunohistochemical staining.STRO-1 was positive in cells by immunofluorescence.Compared with negative control group, the activities of alkaline phosphatase increased after treatment with rhBMP₂ for 7,14 and 21d(P<0.01),the expression of Delta protein was higher after treatment with rhBMP₂ for 21d by westem blotting (P<0.05). Conclusion The cultured human dental pulp stem cells have have the characteristics of stem cells.rhBMP₂ may increase the activity of alkaline phosphatase and up-regulate the expression of. Delta protein may be involved in the differentiation process of Human dental pulp stem cells in to odontoblasts like cells.

Key words: Delta protein , dental pulp stem cells

中图分类号:

R781

于娜,何飞，谭颖徽. rhBMP₂对人牙髓干细胞分化及Delta蛋白表达的影响[J]. J4, 2009, 35(2): 330-333.

YU Na, HE Fei, TAN Ying-hui. Effects of rhBMP₂ on differentiation of human dental pulp stem cells and expression of Delta protein in vitro[J]. J4, 2009, 35(2): 330-333.

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rhBMP₂对人牙髓干细胞分化及Delta蛋白表达的影响

Effects of rhBMP₂ on differentiation of human dental pulp stem cells and expression of Delta protein in vitro

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