PKH-26标记骨髓间充质干细胞气管移植的体内示踪

背景：PKH-26已经被成功应用于标记骨髓间充质干细胞并进行体内示踪。目的：通过气管移植合并静脉移植PKH-26标记的骨髓间充质干细胞评价PKH-26的示踪效果，并检测骨髓间充质干细胞在体内向受损组织的迁移情况。方法：用贴壁法进行骨髓间充质干细胞的原代细胞培养，以PKH-26标记3~5代骨髓间充质干细胞经鼠尾静脉移植入气管移植的受体大鼠体内，用等量的PBS注射作为实验对照。 结果与结论：用PKH-26进行细胞骨髓间充质干细胞标记后，荧光显微镜下观察可见几乎所有骨髓间充质干细胞均有红色荧光。移植后8周仍可见受体气管和移植气管处有红色荧光标记。远离吻合口的受体气管基本未见红色荧光标记物。结果表明骨髓间充质干细胞经静脉移植后，可向损伤的气管组织迁移并定植，而且可随移植气管的血管化逐渐由受体气管切缘向移植气管迁移，最后可定植于移植段气管。PKH-26的红色荧光标记稳定性高，是可应用于长期观察的无降解的荧光标记物。

Labeling and tracing bone marrow mesenchymal stem cells with PKH-26 in vivo

BACKGROUND:PKH-26 has been successfully applied to label bone marrow mesenchymal stem cells (BMSCs). OBJECTIVE:To evaluate the effect of PKH-26 on the labeling and tracing of BMSCs via tracheal transplantation and vein transplantation and to detect the migration of labeled BMSCs into the injured sites.METHODS:BMSCs were cultured by adherence method. PKH-26 labeled 3-5 passage BMSCs were injected into the recipient rats via the tail vein. Rats in the control groups were injected with the same amount of PBS. RESULTS AND CONCLUSION:After PKH-26 labeling procedure, almost all BMSCs were labeled with red fluorescence under inverted fluorescence microscope, indicating that the labeling method has high sensitivity and efficiency. In vivo tracing showed that red fluorescence was still visible at 8 weeks post-surgery at the recipient and transplanted trachea, with most red fluorescence shown at the anastomotic part. There was basically no red fluorescence visible at the recipient trachea that was further away from the anastomotic site. These indicated that BMSCs can migrate to the impaired trachea after injected via the tail vein. Also, the BMSCs can further migrate from the recipient tracheal edge to the transplanted trachea and finally locate at the implant site. The highly stable PKH-26 red fluorescence in vivo indicated that it can be used in tracing studies that need long-term observation.