条件培养液诱导牙龈间充质干细胞分化的体外实验研究

doi:10.3969/j.issn.1005-4979.2015.03.002

《口腔颌面外科杂志》 ›› 2015, Vol. 25 ›› Issue (3): 167-. doi: 10.3969/j.issn.1005-4979.2015.03.002

条件培养液诱导牙龈间充质干细胞分化的体外实验研究

陈岩，冯妍慧芝，刘宏伟

同济大学口腔医学院牙周教研室，口腔生物医学及转化医学实验室，上海 200072

出版日期:2015-06-28 发布日期:2015-12-01
通讯作者: 刘宏伟，教授. E-mail:hwliu@tongji.edu.cn
作者简介:陈岩（1987—），女，吉林省长春市人，硕士. E-mail: cyheihei.1987@163.com
基金资助:
国家自然科学基金(30772418、81271152)；上海市科学技术委员会科研计划(074119514)

Investigation of Multipotent of Gingival Mesenchymal Stem Cells Induced by Apical Tooth Germ Cell-conditioned Medium

CHEN Yan, FENG Yan-huizhi, LIU Hong-wei

Department of Periodontology, Laboratory of Oral Biomedical Science and Translation Medicine, School of Stomatology, Tongji University, Shanghai 200072, China

Online:2015-06-28 Published:2015-12-01

摘要/Abstract

摘要： 目的：建立牙龈间充质干细胞（GMSCs）与根端牙胚条件培养液（APTG-CM）的直接共培养体系，检测其体外分化情况。方法：有限稀释法获得单克隆来源的GMSCs，在体外多向诱导分化，检测ALP活性及基因表达（OCN、BSP、ALP、COL-1、CP23）情况。结果：有限稀释法获得的GMSCs可诱导向成骨、成脂肪方向分化。经APTG-CM诱导后，细胞增殖活性和ALP活性均升高；OCN、BSP、ALP、COL-1、CP23基因表达增高。结论：GMSCs经诱导后具备矿化能力；APTG-CM可诱导GMSCs分化，有利于将GMSCs运用于牙周组织工程的研究。

关键词: 人类, 牙龈间充质干细胞, 增殖, 细胞分化

Abstract: Objective: To establish an indirect co-culture system of rat apical tooth germ-conditioned medium（APTG-CM）and gingival mesenchymal stem cells（GMSCs） for investigating differentiation potential in vitro, in order to find a new seed cell for periodontal tissue engineering. Methods: Using the limiting dilution technique, single-colony derived human GMSCs were isolated and expanded to obtain homogeneous populations. Alkaline phosphatase (ALP) activity, mineralization behavior, gene expression of cementoblast phenotype (OCN, BSP, ALP, COL-1, CP23) were evaluated in vitro. Results: GMSCs were capable of forming colonies and osteogenic and adipogenic differentiations under induction. The induced GMSCs exhibited several characteristics of cementoblast lineages, as indicated by the morphological changes, increased proliferation, high ALP activity and the expression of cementum-related genes in vitro. Conclusion: GMSCs could obtain the ability of mineralization, and apical tooth germ cell-conditioned medium may promote gingival mesenchymal stem cells differentiation.

Key words: human, gingival mesenchymal stem cells(GMSCs), proliferation, differentiation

中图分类号:

R782

陈岩，冯妍慧芝，刘宏伟. 条件培养液诱导牙龈间充质干细胞分化的体外实验研究[J]. 《口腔颌面外科杂志》, 2015, 25(3): 167-.

CHEN Yan, FENG Yan-huizhi, LIU Hong-wei. Investigation of Multipotent of Gingival Mesenchymal Stem Cells Induced by Apical Tooth Germ Cell-conditioned Medium[J]. 《Journal of Oral and Maxillofacial Surgery》, 2015, 25(3): 167-.

https://journal06.magtech.org.cn/Jweb_joms/CN/Y2015/V25/I3/167

参考文献

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[8] Yu JH, Shi JN, Deng ZH, et al. Cell pellets from dental papillae can reexhibit dental morphogenesis and dentinogenesis[J]. Biochem Biophys Res Commun, 2006, 346(1):116-124.

[9] Yang ZH, Zhang XJ, Dang NN, et al. Apical tooth germ cellconditioned medium enhances the differentiation of periodontal ligament stem cells into cementum/periodontal ligament-like tissues[J]. J Periodont Res, 2009, 44(2):199-210.

[10] Tomar GB, Srivastava RK, Gupta N, et al. Human gingiva-derived mesenchymal stem cells are superior to bone marrow-derived mesenchymal stem cells for cell therapy in regenerative medicine[J]. Biochem Biophy Res Commun, 2010, 393(3):377-383.

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[12] Kumar S, Mahendra G, Ponnazhagan S. Determination of osteoprogenitor-specific promoter activity in mouse mesenchymal stem cells by recombinant adeno-associated virus transduction[J]. Biochim Biophys Acta, 2005, 1731(2):95-103.

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条件培养液诱导牙龈间充质干细胞分化的体外实验研究

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条件培养液诱导牙龈间充质干细胞分化的体外实验研究

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