安徽医科大学学报2026，Vol.61Issue(7)：1163-1169,7.DOI:10.19405/j.cnki.issn1000-1492.2026.07.001
流式细胞术分选小鼠肿瘤髓源性抑制细胞的方案优化及鉴定
Optimization and characterization of a protocol for sorting mouse tumor myeloid-derived suppressor cells by flow cytometry
摘要
Abstract
Objective To optimize the flow cytometric sorting workflow for myeloid-derived suppressor cells(MD-SCs)from murine Lewis lung carcinoma(LLC)subcutaneous tumor tissues,improving single-cell suspension preparation efficiency and sorting purity,thereby providing high-quality cells for subsequent functional studies.Methods Subcutaneous LLC tumor tissues were harvested from C57BL/6 mice,and two enzymatic digestion proto-cols were first compared to evaluate their effects on single-cell suspension preparation.After staining with a viabil-ity dye(Live/Dead)and fluorochrome-conjugated anti-mouse antibodies against CD45,CD11b,and Gr-1,cells were loaded onto a Beckman CytoFLEX cell sorter.Next,conventional and optimized gating strategies were ap-plied.In the optimized strategy,the CD45+population was used as the initial gate to directly define an MDSC-enriched region,followed by selection of CD11b+Gr-1+cells.Finally,purity was assessed by flow cytometry,vi-ability was determined by trypan blue staining,and RT-qPCR was performed to measure the expression of MDSC signature genes,including Arg1,Nos2,IL-10,and S100A8/A9.Results The optimized enzymatic digestion proto-col significantly increased the yield and viability of single cells.After optimization of the gating strategy,the post-sort positivity rate(purity)was markedly improved.Arg1,Nos2,IL-10 and S100A8/A9 were highly expressed in the sorted cells,indicating that the isolated cells retained characteristic MDSC features and functional signatures.Conclusion By optimizing both the tissue digestion protocol and the flow cytometric gating strategy,we establish an efficient workflow for isolating MDSCs from tumor tissues.This method improves the yield and purity of single-cell preparations while preserving antigen integrity,providing a reliable methodological foundation for subsequent studies on MDSC function and metabolism.关键词
髓源性抑制细胞/流式细胞术/细胞分选/方案优化/Lewis肺癌/肿瘤微环境Key words
myeloid-derived suppressor cells/flow cytometry/cell sorting/protocol optimization/Lewis lung carcinoma/tumor microenvironment分类
医药卫生引用本文复制引用
杜佳,饶路路,侯昕,马旸..流式细胞术分选小鼠肿瘤髓源性抑制细胞的方案优化及鉴定[J].安徽医科大学学报,2026,61(7):1163-1169,7.基金项目
国家自然科学基金项目(编号:82170603、81502123) （编号:82170603、81502123）
宁波市自然科学基金重点项目(编号:2024J033) National Natural Science Foundation of China(Nos.82170603,81502123) （编号:2024J033）
Key Project of Natural Science Foundation of Ningbo(No.2024J033) （No.2024J033）
