摘要
目的:探讨人脐带间充质干细胞(hUC-MSCs)上清凝胶对小鼠皮肤创面修复的作用,为临床皮肤创伤提供新的修复方法奠定基础。方法:从新生儿脐带分离出hUC-MSCs并培养,经流式细胞仪检测细胞表面标志物、成骨成脂细胞分化实验以及RT-PCR检测干性相关基因的表达证明其干细胞特性;收集上清液,进行粗提纯及总蛋白浓度检测,制成凝胶剂,用于小鼠背部损伤模型中创面的治疗;于第2、3、6、8、15 d取样,伤口及正常皮肤组织样品分别用于制备组织病理切片及愈合组织相关因子表达的Q-PCR检测。结果:从新生儿脐带分离出的hUC-MSCs其形态、表面标志物符合干细胞特性,具有成骨和成脂诱导分化潜能。实验小鼠皮肤创伤后3 d开始可见各组创面逐渐缩小,伤后15 d创面基本愈合;第5、7 d高剂量组创面愈合率大于对照组,其差异有统计学意义(P<0.05)。病理切片结果显示高剂量组伤口愈合程度最高,低剂量组次之,两者都优于对照组。愈合组织相关因子检测表明,伤后第2 d,高剂量上清处理的小鼠伤口组织TGF-β1和Col1a1基因表达增加,明显多于对照组(P<0.05),伤后第3 d,实验组VEGF的表达量均高于对照组(P<0.05);高剂量组从第3 d开始,IL-1β这一炎症因子表达水平显著低于空白对照组(P<0.05)。结论:hUC-MSCs上清凝胶能够提高小鼠皮肤创面愈合的速度。
Objective： To explore the effect of hmnan mnbilical cord mesenehynlal stem cells（hUC-MSCs） supernatant gel on the repair of skin wounds in mice, and to lay a foundation for providing new repair methods for clinical skin wounds. Methods： hUC-MSCs were isolated from the umbilical cord of the newborn and cultured, and their surface markers were detected by flow eytometiT. Then differentiation experiments of adipogenie and oste- obiogenesis cells and RT-PCR detection of stem-related genes were performed to prove their stem cell characteristics. The culture supernatant was collected for crude purification, and total protein concentration was detected. And then it was made into gel for the treatment of wounds in the mouse back injury model. Samples were taken at day 2, 3, 6, 8, and 15. Wound and normal skin tissue samples were used to prepare histopathologieal sections and Q-PCR detection of tissue-related factors. Results： The morphology and surface markers of hUC-MSCs isolated from the umbilical cord of newborns were consistent with the characteristics of stem cells and had the potential of bone and fat induced differentiation. Three days after the skin trauma of the experimental mice, it can be seen that the wounds of each group gradually narrowed. And the wounds healed 15 days after the injury. On the day 5 and 7 the wound healing rate of the high dose group was greater than that of the control group, and the difference was statistically significant（P〈0.05）. The pathological results showed that the wound healing was the best in the high dose group and the second in the low dose group, both of which were superior to the control group. The test of the con＇elative factor fl＇om healing tissue showed that the TGF-fil and Collal gene expression in the high-dose treated mice wound tissue increased significantly on the second day after the injury, significantly more than in the control group（P〈0.05）. The expression of VEGF was higher in the experimental group than in the control group on the day 3 after injury（P〈0.05）. From the day 3, the expression of the inflammatory factor IL-1β was significantly lower in the high dose group than in the control group. Conclusion： hUC-MSCs can improve the rate of skin wound healing in mice.
作者
冷晓燕
段云飞
段颖
徐燕
常菁
LENG Xiao-Yan;DUAN Yun-Fei;DUAN Ying;XU Yan;CHANG Jing(Jiangsu Maijian Biotechnology Development Co.,Ltd.,Wuxi 214000,China)
出处
《生物技术通讯》
CAS
2018年第5期659-666,共8页
Letters in Biotechnology
关键词
人脐带间充质干细胞
干细胞上清
凝胶
伤口愈合
human umbilical cord mesenchymal stem cells（hUC-MSCs）
stem cells culture supernatant
gelata
wound healing
