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 cel transplantation tracer methods require histological analysis and identification in vitro, which limits the clinical application of stem cel 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 cel s from macaques. METHODS:Bone marrow mesenchymal stem cel s were derived from bone marrow aspirates of healthy macaques using whole bone marrow adherence method. Then, the cel s were identified using flow cytometry detection. Bone marrow mesenchymal stem cel s 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 cel viability and proliferation. RESULTS AND CONCLUSION:Bone marrow mesenchymal stem cel s were successful y isolated from healthy macaques using the whole bone marrow adherence method, and the cel purity was up to 95.1%. SPIO and DAPI were both successful to label the bone marrow mesenchymal stem cel s with a positive rate of 95%-98%, but had no influence on cel viability and proliferation.
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