不同供体来源P5代次人脐带间充质干细胞免疫调节能力的差异性研究  

Difference in immune regulation ability of p5 human umbilical cord mesenchymal stem cells from different donor sources

作　　者：吴天韵 邢忠刚 解华晓 张鑫[1] 陈雷[1] 程黎明[1] Wu Tianyun;Xing Zhonggang;Xie Huaxiao;Zhang Xin;Chen Lei;Cheng Liming(Key Laboratory of Spine and Spinal Cord Injury Repair and Regeneration,Shanghai Tongji Hospital,Shanghai 200065,China)

机构地区：[1]上海市同济医院骨科脊柱脊髓损伤再生修复教育部重点实验室,200065

出　　处：《中华实验外科杂志》2022年第4期691-693,共3页Chinese Journal of Experimental Surgery

摘　　要：目的研究不同供体来源的P5代次人脐带间充质干细胞(hUC-MSCs)免疫调节功能的差异性。方法随机选取3株不同供体的hUC-MSCs培养至P5代并检测其表面标志物、细胞周期及分化能力。将P5代hUC-MSCs与外周血单个核细胞(PBMCs)直接接触培养,使用流式细胞仪检测不同供体来源的hUC-MSCs对淋巴细胞亚群及细胞因子的免疫调节效果。收集P5代细胞上清液加入到小鼠小胶质细胞(BV2)的培养体系中检测BV2细胞的存活率。多组数据组间比较采用单因素方差分析。结果不同供体来源的P5代hUC-MSCs实验组中淋巴细胞增殖占比显著低于对照组[(47.47±2.48)%比(23.83±0.96)%,(16.33±0.50)%,(16.23±1.02)%,F=312.96,P<0.01];实验组中Th1细胞亚群的增殖占比显著低于对照组[(5.48±0.20)%比(3.23±0.04)%,(4.49±0.08)%,(3.82±0.05)%,F=143.072,P<0.01];实验组中Th17细胞亚群的增殖占比显著低于对照组[(1.33±0.06)%比(0.85±0.01)%,(1.00±0.01)%,(0.76±0.02)%,F=161.762,P<0.01];实验组中BV2细胞的存活率显著低于对照组[(100.00±0.00)%比(15.92±0.02)%,(12.91±0.011)%,(79.86±0.08)%,F=168.857,P<0.01],差异有统计学意义;实验组中Treg细胞增值占比显著高于对照组[(8.07±0.33)%比(11.47±0.57)%,(11.87±0.64)%,(11.57±0.72)%,F=28.079,P<0.01]和对TNF-α分泌量的抑制效果(360.74±7.98比24.24±0.44,27.90±0.90,34.89±1.90,F=3224.630,P<0.01),差异有统计学意义。结论不同供体来源的P5代次hUC-MSCs均有分化潜能,但免疫调节能力存在差异,在临床应用中应先对hUC-MSCs进行筛选以保证免疫调节能力的有效性。Objective To investigate the differences in immunomodulatory functions of human umbilical cord mesenchymal stem cells(hUC-MSCs)from different donors.Methods The hUC-MSCs of 3 strains were randomly selected and cultured to passage 5(P5),and their surface markers,cell cycle and differentiation potential were determined.Human peripheral blood mononuclear cells were co-cultured with 3 strains of P5 hUC-MSCs.Flow cytometry was used to detect the immunomodulatory effects of huc-MSCs on lymphocyte subsets and cytokines.The supernatant of P5 hUC-MSCs was collected and added into the culture system of BV2 microglial cells.The survival rate of BV2 microglial cells was tested.One-way Anova(S-NK)was used as statistical analysis to compare the quantitative data and P-value<0.05 was considered to be significant difference.Results P5 hUC-MSCs from different donors had a significantly inhibitory effect on lymphocytes[(47.47±2.48)%vs.(23.83±0.96)%,(16.33±0.50)%,(16.23±1.02)%,F=312.96,P<0.01].In addition,P5 huc-MSCs could significantly inhibit both Th1 cell subsets[(5.48±0.20)%vs.(3.23±0.04)%,(4.49±0.08)%,(3.82±0.05)%,F=143.072,P<0.01],and Th17 cell subsets[(1.33±0.06)%vs.(0.85±0.01)%,(1.00±0.01)%,(0.76±0.02)%,F=161.762,P<0.01].Furthermore,huc-MSCs significantly inhibited BV2 cells[(100.00±0.00)%vs.(15.92±0.02)%,(12.91±0.011)%,(79.86±0.08)%,F=168.857,P<0.01].There were significant differences among groups(P<0.01).However,there was no significant difference among groups(P>0.05)in inhibitory effect on TNF-αsecretion[(8.07±0.33)%vs.(11.47±0.57)%,(11.87±0.64)%,(11.57±0.72)%,F=28.079,P<0.01]and the promotion of Treg cell subsets(360.74±7.98 vs.24.24±0.44,27.90±0.90,34.89±1.90,F=3224.630,P<0.01).Conclusion P5 hUC-MSCs had inhibitory effects on TNF-αsecretion,and there was no intergroup difference.Therefore,huc-MSCs should be screened prior to clinical application to ensure the ability of immune regulation.

关 键 词：脐带间充质干细胞 免疫调节 外周血单个核细胞 

分 类 号：R329.2[医药卫生—人体解剖和组织胚胎学]