SPIO、DAPI双重标记猕猴骨髓间充质干细胞：对细胞活性及增殖的影响  

SPIO and DAPI double labeled bone marrow mesenchymal stem cells of macaques: effects on cell viability and proliferation

作　　者：宋巧巧[1] 周慧良 潘兴华[3] 

机构地区：[1]湖北科技学院临床医学院内科教研室,湖北省咸宁市437100 [2]咸宁市中心医院,湖北省咸宁市437100 [3]解放军昆明总医院干细胞与组织器官工程研究中心,云南省干细胞工程室验室,云南省昆明市650032

出　　处：《中国组织工程研究》2015年第36期5741-5745,共5页Chinese Journal of Tissue Engineering Research

基　　金：国家自然科学基金资助项目(31172170)~~

摘　　要：背景:传统的细胞移植示踪方法均需要在体外状态下进行组织学切片鉴定和分析,这显然限制了干细胞移植进入临床应用,因此,探索无创的、可多次重复应用的活体示踪方法已成为迫切亟待解决的问题。目的:观察SPIO和DAPI双标记猕猴骨髓间充质干细胞的效果及对干细胞存活、增殖的影响。方法:全骨髓贴壁法分离培养猕猴骨髓间充质干细胞,并采用流式细胞术对骨髓间充质干细胞进行鉴定。用SPIO和DAPI双标记骨髓间充质干细胞,通过荧光显微镜观察DAPI标记阳性率,普鲁士蓝染色和透射电镜鉴定SPIO标记阳性率,MTT检测标记细胞的活性、增殖能力。结果与结论:采用全骨髓贴壁法成功获得高纯度猕猴骨髓间充质干细胞,纯度达95.1%。SPIO和DAPI成功标记骨髓间充质干细胞,标记阳性率达95%-98%,且对干细胞的活性、增殖无影响。BACKGROUND： Traditional cell transplantation tracer methods require histological analysis and identification in vitro, which limits the clinical application of stem cell transplantation. So it is urgent to establish an in vivo noninvasive and repeatable tracer method. OBJECTIVE： To observe the effect of SPIO and DAPI double labeling on survival and proliferation of bone marrow mesenchymal stem cells from macaques. METHODS： Bone marrow mesenchymal stem cells were derived from bone marrow aspirates of healthy macaques using whole bone marrow adherence method. Then, the cells were identified using flow cytometry detection. Bone marrow mesenchymal stem cells were labeled using SPIO and DAPI. Fluorescent microscope was used to detect DAPI positive rate, and Prussian blue staining and transmission electron microscope were employed to measure SPIO positive rate. MTT assay was used to detect cell viability and proliferation. RESULTS AND CONCLUSION： Bone marrow mesenchymal stem cells were successfully isolated from healthy macaques using the whole bone marrow adherence method, and the cell purity was up to 95.1%. SPIO and DAPI were both successful to label the bone marrow mesenchymal stem cells with a positive rate of 95%-98%, but had no influence on cell viability and proliferation.

关 键 词：干细胞 骨髓干细胞 细胞标记 骨髓间充质干细胞 猕猴 SPIO DAPI 国家自然科学基金 

分 类 号：R394.2[医药卫生—医学遗传学]