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目的研究不同供者来源的人脐带间充质干细胞(h UC-MSC)分泌的外泌体免疫调节能力的异质性。方法培养5株不同供者来源的h UC-MSC,收集无血清培养上清提取外泌体,流式细胞术检测其表面特异性标志CD9、CD63、CD81、CD44的表达,BCA法测定总蛋白含量。将外泌体和脐带血单个核细胞(UBMC)共培养72 h,MTT实验检测细胞增殖活性,ELISA测定共培养上清中干扰素γ(IFN-γ)和肿瘤坏死因子α(TNF-α)的含量。离心重悬UBMC和K562细胞按5∶1的比例共培养4 h,MTT实验检测细胞活性,计算UBMC的杀伤能力。结果提取的外泌体表达CD9,CD63,CD81及CD44。不同供者来源h UC-MSC分泌的外泌体对UBMC的增殖、分泌细胞因子及K562细胞杀伤能力的作用不同。结论不同供者h UC-MSC的外泌体免疫调节能力具有异质性。
Objective To investigate the heterogeneity of immunoregulatory capacity of exosomes secreted by human umbilical cord mesenchymal stem cells (h UC-MSCs) from different donor sources. Methods Five hUC-MSCs from different donors were cultured, and the supernatant of serum-free medium was collected to extract exosomes. The expression of CD9, CD63, CD81 and CD44 were detected by flow cytometry. Total protein content. The exosomes and umbilical cord blood mononuclear cells (UBMC) were co-cultured for 72 h. The cell proliferation activity was detected by MTT assay. The levels of IFN-γ and TNF-α in co-culture supernatants were determined by ELISA. Content. Centrifuge resuspended UBMC and K562 cells were co-cultured at a ratio of 5: 1 for 4 h. The activity of UBMC was calculated by MTT assay. Results The exosomes extracted expressed CD9, CD63, CD81 and CD44. The exosomes secreted by h UC-MSCs from different donor sources have different effects on the proliferation and secretion of cytokines and the cytotoxicity of K562 cells. Conclusion The exosomes of h UC-MSCs from different donors have heterogeneous immunomodulatory capacity.