PO.TB10.02 · 肿瘤生物学
开发用于脑肿瘤模型中小胶质细胞和免疫细胞群体的流式细胞术检测panel
Development of a flow cytometry panel for microglia and immune cell populations in a brain tumor model
作者与单位 Authors & Affiliations
摘要 Abstract
中文摘要
准确表征小胶质细胞和浸润性免疫细胞群体对于理解脑肿瘤的免疫学格局以及评估治疗反应至关重要。在此，我们报告了一种多色流式细胞术panel的开发与优化，该panel专门用于在实验性脑肿瘤模型中区分驻留小胶质细胞与外周髓系和淋巴系亚群。标志物选择以组织特异性表达谱为指导，利用CD45、CX3CR1和P2RY12的差异水平来分辨小胶质细胞，同时纳入浸润性巨噬细胞、树突状细胞、中性粒细胞和T细胞的经典标志物。对酶解条件、活力染料选择和门控层次进行了系统优化，以保持表位完整性并最大化信号分辨率。使用荷瘤和对照脑组织进行的验证证明，小胶质细胞群体与浸润性群体得到稳健的分离，且在各生物学重复间具有可重复的定量。该工作流程为脑肿瘤研究中的免疫表征提供了一种可靠且可扩展的方法，从而能够更深入地洞察神经免疫相互作用以及免疫治疗驱动的肿瘤微环境内变化。
查看英文原文 English abstract
Accurate characterization of microglia and infiltrating immune cell populations is essential for understanding the immunological landscape of brain tumors and for evaluating therapeutic responses. Here, we report the development and optimization of a multicolor flow cytometry panel tailored to discriminate resident microglia from peripheral myeloid and lymphoid subsets within an experimental brain tumor model. Marker selection was guided by tissue-specific expression profiles, leveraging differential levels of CD45, CX3CR1, and P2RY12 to resolve microglia, while incorporating canonical markers for infiltrating macrophages, dendritic cells, neutrophils, and T cells. Enzymatic dissociation conditions, viability dye selection, and gating hierarchy were systematically optimized to preserve epitope integrity and maximize signal resolution. Validation using tumor-bearing and control brain tissues demonstrated robust separation of microglial and infiltrating populations with reproducible quantification across biological replicates. This workflow provides a reliable and scalable approach for immune profiling in brain tumor research, enabling deeper insight into neuroimmune interactions and immunotherapy-driven changes within the tumor microenvironment.
利益披露 Disclosure
H. Ketteringham, None..
C. Davis, None.