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    人脐带间充质干细胞治疗糖尿病创面效果及其在体内存活、定植研究

    Effects of human umbilical cord mesenchymal stem cells in the treatment of diabetic wound, survival and colonization in vivo

    • 摘要: 目的 探讨人脐带来源间充质干细胞(human umbilical cord mesenchymal stem cells, hUC-MSCs)治疗裸鼠糖尿病创面效果,并观察其在体内存活以及定植分化情况。方法 采用腹腔注射STZ法诱导糖尿病裸鼠,然后于裸鼠背部切取约1.5 cm×1.5 cm的创面。CM-Dil荧光染料处理hUC-MSCs并移植于裸鼠创面;形态学观察并计算创面愈合率;动物活体成像仪检测注射0 h、12 h、1 d、2 d、7 d、14 d后裸鼠创面hUC-MSCs荧光通量;相应时间点取裸鼠创面基底及创缘皮肤,切片后荧光显微镜下观察分析皮肤中移植细胞的分布和存活情况;注射后第14 d取出裸鼠心脏、肝脏、脾脏、肺脏、肾脏、脑等组织,荧光显微镜下观察hUC-MSCs分布情况。结果 与对照组相比,hUC-MSCs可显著促进创面愈合;注射移植hUC-MSCs后,小动物活体成像仪显示染色后hUC-MSCs的荧光集中分布在创面周围并向创面基底迁移,且在移植后24~72 h浓度最高,此后细胞数量迅速下降;移植14 d后,动物体内未见移植的hUC-MSCs存活。结论 hUC-MSCs可促进裸鼠创面愈合,其在移植后24~72 h局部浓度最高,但不能在动物体内长期定植分化。

       

      Abstract: Objective To investigate the effects of human umbilical cord mesenchymal stem cells (hUC-MSCs) in the treatment of diabetic wounds in nude mice, and to observe the survival, colonization and differentiation of hUC-MSCs. Methods Diabetic nude mice were induced by intraperitoneal injection of STZ, and a 1.5 cm×1.5 cm diameter wound was cut from the back of the nude mice. Then, hUC-MSCs were treated with CM-Dil stain and transplanted into the wound of nude mice. The morphological changes were observed to calculate the wound healing rate. The fluorescence flux of hUC-MSCs on the wound surface of nude mice after injection after 0 h, 12 h, 1 day, 2 days, 7 days and 14 days was measured by the small animal live imaging system. The skin at the base and edge of the wound of the nude mice were taken at the corresponding time points, and the distribution and survival of the transplanted cells in the skin were observed under a fluorescence microscope. On the 14th day after injection, the heart, liver, spleen, lungs, kidneys, brain and other tissues of the nude mice were taken, and the distribution of hUC-MSCs was observed under a fluorescence microscope. Results Compared with the control group, hUC-MSCs significantly promote wound healing. After hUC-MSC transplantion, the small animal live imaging system showed that the fluorescence of stained hUC-MSCs was concentrated around the wound surface and migrated to the base of the wound surface. The concentration was the highest 24—72 h after transplantation, and then the number of the cells rapidly decreased. After 14 days of transplantation, no hUC-MSCs were found to survive within the mice. Conclusions hUC-MSCs can promote wound healing in nude mice, and the local concentration of hUC-MSCs is the highest 24—72 h after transplantation. However, hUC-MSCs cannot colonize and differentiate in animals for a long time.

       

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