论文部分内容阅读
目的研究过表达CC趋化因子受体7(CCR7)的C3H10 T1/2小鼠间充质干细胞系在小鼠皮肤移植模型中,是否具有比原始C3H10 T1/2细胞更强的免疫抑制作用。方法培养扩增稳定的小鼠C3H10 T1/2细胞,并利用慢病毒工具导入EGFP-CCR7基因;以C57BL/6小鼠为供体获取皮片,以BALB/c小鼠为受体建立小鼠同种异系皮肤移植模型;实验动物分4组:CCR7+C3H10 T1/2细胞组、C3H10 T1/2组、同种异系对照组和同系对照组(受者小鼠移植同系小鼠皮片),分别于移植模型成功当天,经尾静脉注射1×106C3H10 T1/2细胞、1×106CCR7+C3H10 T1/2细胞、等体积生理盐水(对照组);观察皮片移植后的存活情况,并于皮片移植第12天,测定小鼠脾淋巴细胞中Th17细胞与调节性T细胞(Treg)的比例,HE染色观察皮片病理变化。结果受者小鼠皮片移植后12 d,外观、组织病理形态观察以及流式细胞术检测结果表明,CCR7+C3H10 T1/2细胞与C3H10 T1/2细胞的皮片存活情况均优于同种异系对照组;CCR7+C3H10 T1/2细胞皮片移植后存活情况优于C3H10 T1/2细胞组,且移植免疫反应程度轻于C3H10 T1/2细胞组。结论 CCR7+C3H10 T1/2细胞相对于正常的C3H10 T1/2细胞,拥有更好的免疫抑制效应,可有效的降低移植免疫炎性反应,改善移植物的生存状态。
Objective To investigate whether C3H10 T1 / 2 mouse mesenchymal stem cells that over-express CC chemokine receptor 7 (CCR7) have a stronger immunosuppressive effect than the original C3H10 T1 / 2 cells in a mouse skin graft model. Methods The stable C3H10 T1 / 2 cells were expanded and transfected with EGFP-CCR7 gene by lentivirus. C57BL / 6 mice were used as skin grafts and BALB / c mice were used as recipients Allograft skin graft model was divided into 4 groups: CCR7 + C3H10 T1 / 2 cell group, C3H10 T1 / 2 group, allograft control group and homologous control group ), 1 × 106C3H10 T1 / 2 cells and 1 × 106CCR7 + C3H10 T1 / 2 cells were injected into the caudal vein on the day of successful transplantation. The same volume of saline (control group) was injected into the tail vein. On day 12 of skin grafting, the ratio of Th17 cells to regulatory T cells (Tregs) in splenic lymphocytes of mice was measured. Pathological changes of the skin were observed by HE staining. Results The appearance, histopathology and flow cytometry results showed that the survival of the skin of CCR7 + C3H10 T1 / 2 and C3H10 T1 / 2 cells was better than that of the same The survival rate of CCR7 + C3H10 T1 / 2 cell skin graft was better than that of C3H10 T1 / 2 cell group, and the degree of graft immunoreaction was lower than that of C3H10 T1 / 2 cell group. Conclusion Compared with normal C3H10 T1 / 2 cells, CCR7 + C3H10 T1 / 2 cells have better immunosuppressive effect, which can effectively reduce the immunological inflammatory reaction and improve the survival of the grafts.