用BODIPY染色检测小鼠乳腺肿瘤骨髓源性抑制细胞中性脂的方案。

IF 1.3 Q4 BIOCHEMICAL RESEARCH METHODS

STAR Protocols Pub Date : 2025-01-03 DOI:10.1016/j.xpro.2024.103485

Ukjin Kim, Rumela Chakrabarti

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引用次数: 0

摘要

中性脂影响髓源性抑制细胞（MDSCs）的免疫抑制功能。在这里，我们提出了一种使用来自三阴性乳腺癌细胞4T1的小鼠乳腺肿瘤的BODIPY测量MDSCs中中性脂质的方案，该方案适用于其他感兴趣的乳腺肿瘤。我们描述了4T1细胞培养，肿瘤单细胞分离，抗体和BODIPY染色细胞和流式细胞术的步骤。此外，我们引入了MDSC分选的替代方案，以克服BODIPY的细胞死亡风险。有关该协议的使用和执行的完整细节，请参阅Kim等人1。

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Protocol to detect neutral lipids with BODIPY staining in myeloid-derived suppressor cells in mouse mammary tumors.

Neutral lipids affect the immunosuppressive function of myeloid-derived suppressor cells (MDSCs). Here, we present a protocol for measuring neutral lipids in MDSCs using BODIPY from mouse mammary tumor derived from triple-negative breast cancer cells, 4T1, which is applicable to other mammary tumors of interest. We describe steps for 4T1 cell culture, single-cell isolation from tumors, staining of cells with antibodies and BODIPY, and flow cytometry. Furthermore, we introduce alternative protocols with MDSC sorting to overcome risk of cell death by BODIPY. For complete details on the use and execution of this protocol, please refer to Kim et al.¹.

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