摘要
目的:探讨经5-溴脱氧尿嘧啶(5-BrdU)标记的骨髓间充质干细胞(BMSCs)在大鼠创面愈合中的作用。方法:分离培养大鼠BMSCs并用流式细胞仪技术鉴定。将大鼠背部脊柱一侧作为实验组并制作一全层皮肤缺损模型,其对侧为对照组,经尾静脉注入经5-BrdU标记的BMSCs,移植细胞数为2×107个。术后3天、7天、14天、21天、28天分批处死动物,观察移植细胞参与创面愈合的情况。结果:流式细胞仪鉴定细胞表达CD29、CD90表达阳性;CD45表达呈阴性。实验组:经5-BrdU体外标记的BMSCs进行细胞移植14天后大鼠创面局部及边缘可见5-BrdU标记的阳性细胞聚集,免疫组化染色可见基底层、真皮层、新生血管周围有散在红棕色标记细胞;对照组中未发现明显的聚集现象;抗5-BrdU/FⅧ、5-BrdU/波形蛋白免疫双标技术检测结果显示:在创伤部位有部分BMSCs分化为血管内皮细胞和成纤维细胞参与创面愈合。大鼠对侧正常皮肤中未见明显的BMSCs聚集和增殖分化情况。结论:5-BrdU标记的BMSCs参与皮肤创面愈合。
Objective To investigate the contribution of bone marrow mesenchymal stem cells （BMSCs） to skin wound repair, and provide a new thought for mechanism study of wound healing and clinical treatment in the future. Methods Bone marrow-derived BMSCs of SD rats were isolated and cultured in vitro routinely and the fifth passage as taken for identification of specific surface antigens by flow cytometry. Two sides of rats＂ back were randomly derived into two groups, skin resection wounds were produced in the experient group. Labled 5-BrdU BMSCs were implanted through tail vein to model rats. At 3, 7,14,21 and 28 days after BMSCs transplantation, immunohistochemical and immunofluorescence examinations were used to investigate BMSCs aggregate around the wound site and identify the contribution of bone marrow-derived cells to skin wound repair in diffferent time during wound healing. Results Flow cytometry showed that BMSCs expressed CD29, and CD90, but no CD45. At 1,2,3,4 weeks after the cells transplantation, the implanted cells were detected in the wound site in the model rats,which is not founded in normal skin of the rats＇. Part of the implanted cells were detected to differentiate into vascular endothelia cells and fibroblast, while there were not found in the normal skin through BrdU/FⅧ,BrdU/Vimentin double-labelling technique of immunofluorescence. Conclusion Labled 5-BrdU BMSCs contribute to skin wound healing.
出处
《中国美容医学》
CAS
2008年第4期527-530,共4页
Chinese Journal of Aesthetic Medicine
