炎症因子预处理的脂肪干细胞可明显抑制外周血单个核细胞增殖

Inflammatory priming adipose derived stem cells significantly inhibit the proliferation of peripheral blood mononuclear cells

目的:观察炎症因子预处理脂肪干细胞(adipose derived stem cells,ASCs)模拟异常炎症环境能否增强ASCs抑制外周血单个核细胞(peripheral blood mononuclear cells,PBMCs)体外增殖的能力。方法:先用酶消化法对ASCs进行分离及体外培养,流式细胞仪检测其表面CD分子表达情况,用特定培养基诱导分化并鉴定其向脂肪组织和骨组织分化的能力。将PBMCs用CFSE标记后,与炎症因子预处理后的ASCs共培养,对照组无ASCs或使用未经炎症因子处理的ASCs,同时用CD3/CD28刺激PBMCs增殖,最后用流式细胞仪检测PBMC的母代细胞比例,从而判断其增殖情况。结果:ASCs形态为梭形,密集时可成鱼群状生长,可在诱导培养基培养下向脂肪组织和骨组织分化,表面CD分子表达情况与国际脂肪治疗联盟的关于ASCs的说明一致。PBMCs与ASCs体外共培养后,与无ASCs组相比,PBMCs的母代细胞比例有所增加,但效果并不显著,而将PBMCs与用炎症因子预处理后的ASCs共培养,PBMCs的母代细胞比例有明显增加(炎症因子处理后ASCs组38.7%±10%vs.无ASCs组28.4%±8.9%,P<0.05),说明炎症因子预处理的ASCs可明显抑制PBMCs的增殖。结论:炎症因子可增强ASCs的抗炎能力,该发现有助于ASCs更好地在组织修复领域发挥治疗作用。

Objective： To explored whether adipose derived stem cells（ASCs） could inhibit the proliferation of peripheral blood mononuclear cells（PBMCs） and whether inflammatory priming could enhance this property of ASCs. Methods： We isolated ASCs using collagenase from adipose tissue and expanded them in vitro. Cells were induced to differentiate into adipogenic and osteogenic lineages. The cells at passage 3 to passage 5 were used for the experiments. After carboxy fluoresce in succinimidyl ester（CFSE） staining,PBMCs were co-cultured with inflammatory priming ASCs. The PBMCs cultured without ASCs or with non-treated ASCs defined as control groups. Then we used flow cytometry to detect the proliferation of PBMCs. Results： ASCs had fibroblast-like phenotype and were spindle shaped. They were able to differentiate into cells of adipogenic and osteogenic lineages in specific induction media.ASCs had the CD expression profile consistant with the International Federation for Adipose Therapeutics statement. The percentage of parent cells in PBMC after co-cultured with ASCs increased,though there was no statistical significance. However,when co-cultured with inflammatory priming ASCs,the percentage of parent cells significantly increased（with inflammatory priming ASCs group vs. without ASCs group,38. 7% ± 10. 0% vs. 28. 4% ± 8. 9%,P〈0. 05）. This indicated that inflammatory priming ASCs could significantly inhibit the proliferation of PBMCs. Conclusion： Inflammatory cytokines can enhance the immunosuppressive ability of ASCs. Our findings may help the application of ASCs in tissue repairment with better results.