犬牙髓干细胞的分离培养及其生物学特性研究

Isolation,Identification and Biological Characteristics of Dog Dental Pulp Stem Cells

目的:体外分离、培养犬牙髓干细胞,并初步探讨其生物学特性。方法:采用克隆化传代法培养犬牙髓干细胞,获得多克隆来源细胞,检测其克隆形成率,采用流式法鉴定干细胞表面标志物;扩大培养后测定细胞增殖能力及多向分化能力。结果:克隆化传代法可获得呈集落生长的犬牙髓干细胞,其克隆形成率为20%～35%;流式鉴定显示STRO-1、CD34及CD45阳性率分别为14.7%、0.7%、0.8%;犬牙髓干细胞增殖快,增殖曲线呈"S"形;成骨、成脂诱导后碱性磷酸酶、茜素红、油红O染色阳性。结论:克隆化分离培养的犬牙髓干细胞具有间充质干细胞表型和较强的自我更新及多向分化潜能。

Objective： To isolate, culture and explore biological characteristics and potential of multi-directional differentiation of dog dental pulp stem ceils in vitro. Methods： The dog dental pulp stem cells were cultured by cloning subculture, and then expanded for later measurement of biological characteristics. Results： Colony-like dog dental pulp stem cells were available by colony formation assay, and the colony-forming efficiency was 20% -35%. MTT assay indicated high proliferation of dog dental pulp stem cells. Flow cytometry analysis showed that STRO-1 expressed positively, while CD34 and CD45 negatively. Adipogenic and osteogenic differention abilities of dog dental pulp stems cells were confirmed after being induced. Conclusion.. The evidence suggests that dog dental pulp stem cells can be defined as mesenchymal--type cells with the capacity of self-renew and multiple differentiation.