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＝3 224.630， 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。
ObjectiveTo investigate the differences in immunomodulatory functions of human umbilical cord mesenchymal stem cells (hUC-MSCs) from different donors.
MethodsThe 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.
ResultsP5 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-α secretion [(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=3 224.630, P<0.01).
ConclusionP5 hUC-MSCs had inhibitory effects on TNF-α secretion, and there was no intergroup difference. Therefore, huc-MSCs should be screened prior to clinical application to ensure the ability of immune regulation.
吴天韵,邢忠刚,解华晓,等. 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[J]. 中华实验外科杂志,2022,39(04)：691-693.
DOI:10.3760/cma.j.cn421213-20210825-00656版权归中华医学会所有。
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