Ming Liu1, Jinsong Wang, Muyun Liu, Xiang Hu2, Jun Xu3. 1. 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. 2. Shenzhen Beike Cell Engineering Research Institute, Shenzhen 518057, China, Email: huxiang@beike.cc. 3. Email: xufeili@vip.163.com.
Abstract
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.
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.
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