条件培养液诱导牙龈间充质干细胞分化的体外实验研究

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

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。

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, mineral-ization 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, in-creased 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.