PKH26和DAPI联合示踪骨髓干细胞在脑缺血模型小鼠脑内迁移的实验研究

Study on the combination of PKH26 and DAPI to trace bone marrow stem cells homing in mice model of cerebral ischemia

目的探讨PKH26和DAPI联合标记骨髓干细胞在脑内迁移过程中的示踪作用。方法用PKH26和DAPI联合标记从Balb/c小鼠骨髓中分离出的骨髓单个核细胞(BMMCs),用改良电凝法制备局灶性脑缺血模型后,将标记的BMMCs经尾静脉移植入脑缺血小鼠体内,对照组经尾静脉注入PBS。移植2w后取脑组织,通过激光共聚焦荧光显微镜观察BMMCs向脑内迁移的情况。结果实验组小鼠脑组织内可发现PKH26和DAPI双标的神经样细胞。结论 PKH26和DAPI可以联合标记向脑缺血模型小鼠脑组织迁移的骨髓干细胞,为体内示踪骨髓干细胞脑内迁移提供更加准确的实验方法。

Objective To explore the method to trace bone marrow stem cells in mice model of cerebral ischemia by using PKH26 and DAPI fluorescence dye.Methods BMMCs were harvested from BALB/c mice and then doubly labeled with PKH26 and DAPI.The labeled cells were subsequently infused into the tail veins of mice in which focal cerebral ischemia model had been induced by modified electric coagulation approach.14 days after transplantation,the migration status of the cells was studied by fluorescence microscopy.Results Positive cells pre-labeled by PKH26 and DAPI were found in the cortex region of ischemic hemisphere.Conclusions The combined use of PKH26 and DAPI to double label bone marrow stem cells can trace the transplanted cells in mice model of ischemic brain tissues.