人脐带间充质干细胞来源外泌体的免疫调节功能研究
[Study of immunomodulatory function of exosomes derived from human umbilical cord mesenchymal stem cells].
作者信息
Liu Ming, Wang Jinsong, Liu Muyun, Hu Xiang, Xu Jun
机构信息
State Key Laboratory of Respiratory Diseases, Guangzhou Institute of Respiratory Diseases, the First Affiliated Hospital of Guangzhou Medical University, Guangzhou Medical University, Guangzhou 510120, China.
Shenzhen Beike Cell Engineering Research Institute, Shenzhen 518057, China, Email:
出版信息
Zhonghua Yi Xue Za Zhi. 2015 Aug 25;95(32):2630-3.
OBJECTIVE
To investigate the immumodulation ability of exosomes secreted from human umbilical cord-derived mesenchymal stem cells (hUC-MSCs).
METHODS
hUC-MSCs were isolated and cultured.Exosomes were isolated from the culture media of the third-generation hUC-MSCs. The expression of specific surface marker CD9 and CD81 were detected by Western blot, and the concentration of hUC-MSCs exosomes(hUC-MSCs-ex) was evaluated by BCA assay. CD3/CD28-stimulated peripheral blood mononuclear cells(PBMCs) from healthy donor were co-cultured with different concentration of hUC-MSCs-ex for 72 h. The percentage of Th17 and Treg cells and the proliferation of CD4⁺ and CD8⁺ T cells were detected by flow cytometry. ELISA was used to test the level of IFN-γ, IL-6, TNF-α and TGF-β1.
RESULTS
hUC-MSCs-ex inhibited the proliferation of CD4⁺ and CD8⁺ cells obviously,and increased the proportion of CD4⁺ CD25⁺ FoxP3⁺ Treg cells, with high expression of CD81 and CD9. After CD3/CD28 monoclonal antibody stimulated, the percentage of CD45⁺ CD4⁺ Ki67⁺ and CD45⁺ CD8⁺ Ki67⁺ cells were 85.3% ± 5.6% and 72.6% ± 6.3%, respectively. Meanwhile, the level of TGF-β1 were elevated and the level of IFN-γ, IL-6 and TNF-α were decreased (P<0.05).
CONCLUSION
hUC-MSCs-ex has the immunomodulatory functionin vitro, which could be a new therapeutic agent for the treatment of immune disorders.
目的
研究人脐带间充质干细胞（hUC-MSCs）分泌的外泌体的免疫调节能力。
方法
分离培养hUC-MSCs。从第三代hUC-MSCs的培养基中分离外泌体。通过蛋白质免疫印迹法检测特异性表面标志物CD9和CD81的表达，通过BCA法评估hUC-MSCs外泌体（hUC-MSCs-ex）的浓度。将健康供体的经CD3/CD28刺激的外周血单个核细胞（PBMCs）与不同浓度的hUC-MSCs-ex共培养72小时。通过流式细胞术检测Th17和Treg细胞的百分比以及CD4⁺和CD8⁺T细胞的增殖情况。采用酶联免疫吸附测定法检测IFN-γ、IL-6、TNF-α和TGF-β1的水平。
结果
hUC-MSCs-ex明显抑制CD4⁺和CD8⁺细胞的增殖，增加CD4⁺CD25⁺FoxP3⁺Treg细胞的比例，且CD81和CD9高表达。经CD3/CD28单克隆抗体刺激后，CD45⁺CD4⁺Ki67⁺和CD45⁺CD8⁺Ki67⁺细胞的百分比分别为85.3%±5.6%和72.6%±6.3%。同时，TGF-β1水平升高，IFN-γ、IL-6和TNF-α水平降低（P<0.05）。
结论
hUC-MSCs-ex在体外具有免疫调节功能，可能成为治疗免疫紊乱的新型治疗药物。
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