A novel methodology of the myeloid-derived suppressor cells (MDSCs) generation with splenic stroma feeder cells.
A novel methodology of the myeloid-derived suppressor cells (MDSCs) generation with splenic stroma feeder cells.
Exp Cell Res. 2020 May 30;:112119
Authors: Tian F, Jiang P, Wu M, Xing Y, Liu L, Dai X, Tang H, Yang ZF, Wang L, Zhu X
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 pr...
Source: Experimental Cell Research - Category: Cytology Authors: Tian F, Jiang P, Wu M, Xing Y, Liu L, Dai X, Tang H, Yang ZF, Wang L, Zhu X Tags: Exp Cell Res Source Type: research
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