高龄老年人牙髓干细胞分离培养与生物学特性分析实例报告

The preliminary research on geriatric dental pulp stem cells from an 88-year-old man

目的:对高龄老年人牙髓干细胞进行分离培养,探讨其生物学特性。方法:应用联合酶消化法,分离培养一例88岁老年男性患者右上第三磨牙的牙髓细胞,进行原代培养并传代,流式分析鉴定,体外培养观察细胞的形态、增殖、分化能力。结果:首次成功分离培养高龄老年人牙髓干细胞;倒置相差显微镜下可见细胞透亮,呈长梭形;流式细胞学分析显示,体外培养的老年人牙髓干细胞阳性表达间充质干细胞标记CD73、CD90、CD105,不表达造血干细胞和内皮细胞标记CD14、CD19、CD34、CD45及H LA-DR;细胞活力良好,可成骨向分化。结论:高龄老年人牙髓组织中存在牙髓干细胞,活力良好,具有一定的分化潜能。

Objective： To isolate and obtain dental pulp stem cells （DPSCs） from the oldest-old and explore their biological functions. Methods： The dental pulp cells from an 88-year-old male patient＇s upper third molar were isolated using the enzymatic digestion of pulp tissue method, cultured, passaged and analyzed by flow cytometry. Cell morphology, proliferation and differentiation ability were observed in vitro. Results： It is the first to successfully isolated, cultured and passaged the geriatric dental pulp stem cells（GDPSCs）. The morphology of the cells were spindle or exhibited several pseudopods under inverted phase contrast microscope and were positive for human multipotent mesenchymal stromal cells （MSCs） surface markers CD73, CD90, and CD105, but negative for CD14, CD19, CD34, CD45 and HLA-DR. These cells also exhibited good self-renewal ability and multilineage differentiation potential. Conclusion： DPSCs can be isolated from the pulp tissues of the oldest-old and present well proliferation and differentiation potentials.