1. A novel methodology of the myeloid-derived suppressor cells (MDSCs) generation with splenic stroma feeder cells
- Author
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Li-xin Wang, Min Wu, Li Liu, Xinbin Dai, Yun Xing, Fang Tian, Zhong-Fa Yang, Xuejun Zhu, Pengjun Jiang, and Hua Tang
- Subjects
0301 basic medicine ,Male ,Myeloid ,Lymphoma ,Cell Culture Techniques ,Graft vs Host Disease ,Biology ,CD8-Positive T-Lymphocytes ,T-Lymphocytes, Regulatory ,Immunophenotyping ,Colony-Forming Units Assay ,03 medical and health sciences ,Mice ,0302 clinical medicine ,Stroma ,In vivo ,medicine ,Animals ,Cell Proliferation ,Arginase ,Myeloid-Derived Suppressor Cells ,Feeder Cells ,Cell Biology ,Colony-stimulating factor ,Survival Analysis ,030104 developmental biology ,medicine.anatomical_structure ,030220 oncology & carcinogenesis ,Myeloid-derived Suppressor Cell ,Cancer research ,Female ,Bone marrow ,Stromal Cells ,Reactive Oxygen Species ,CD8 ,Spleen - Abstract
Myeloid-derived suppressor cells (MDSCs) are a significant obstacle for immunotherapy of cancer. It is of great clinical relevance to study the mechanism of MDSCs accumulation in mouse spleens and establish a stable method to obtain high-purity MDSCs in vitro for further research. Here, we established a new method for amplifying a large number of highly pure MDSCs in vitro. To mimic the microenvironment of MDSCs development in vivo, mouse splenic stroma feeder cells and serum-free medium containing granulocyte-macrophage colony stimulating factor (GM-CSF) were used to induce myeloid precursors in mouse bone marrow cells, which differentiate into MDSCs. Development and immunological functions of the cells were monitored both in vivo and in vitro. A total of 4 × 108 MDSCs could be obtained from the bone marrow from one mouse, the ratio of CD11b+Gr-1+ MDSCs could reach 93.8% ± 3.3% after nine days of culture in vitro. Cultured MDSCs maintained a similar immunophenotype with MDSCs found in tumor-bearing mice. Colony forming assay in vitro and in vivo demonstrated that these were myeloid precursor cells. These cells generated high levels of reactive oxygen species and arginase 1 to prevent proliferation of CD8+ T cells in vitro. These also increased regulatory T (Treg) cells in blood while promoting the growth of lymphoma in vivo. In addition, cultured MDSCs effectively inhibited acute graft-versus-host disease (aGVHD). Our findings suggest that mouse splenic stroma plays an important role in the generation of MDSCs and represent a preliminary mechanism for the accumulation of MDSCs in spleens, and thereby lay the foundation for basic research and the clinical application of MDSCs.
- Published
- 2020