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在小鼠乳腺肿瘤的髓源性抑制细胞中用BODIPY染色检测中性脂质的实验方案。

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

作者信息

Kim Ukjin, Chakrabarti Rumela

机构信息

Department of Surgery, Sylvester Comprehensive Cancer, University of Miami Miller School of Medicine, Miami, FL 33136, USA.

Department of Surgery, Sylvester Comprehensive Cancer, University of Miami Miller School of Medicine, Miami, FL 33136, USA.

出版信息

STAR Protoc. 2025 Mar 21;6(1):103485. doi: 10.1016/j.xpro.2024.103485. Epub 2025 Jan 3.

DOI:10.1016/j.xpro.2024.103485
PMID:39754723
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11754504/
Abstract

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..

摘要

中性脂质影响髓源性抑制细胞（MDSCs）的免疫抑制功能。在此，我们展示了一种使用BODIPY测量源自三阴性乳腺癌细胞4T1的小鼠乳腺肿瘤中MDSCs中性脂质的方法，该方法适用于其他感兴趣的乳腺肿瘤。我们描述了4T1细胞培养、从肿瘤中分离单细胞、用抗体和BODIPY对细胞进行染色以及流式细胞术的步骤。此外，我们介绍了通过MDSC分选的替代方法，以克服BODIPY导致细胞死亡的风险。有关本方法的使用和执行的完整详细信息，请参考Kim等人的研究。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cf6/11754504/8a7cd6ea3e02/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cf6/11754504/a973aa079ccf/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cf6/11754504/03a95a01d59f/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cf6/11754504/7d3a124ea6db/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cf6/11754504/183218e508ce/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cf6/11754504/1a0f609c0ab9/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cf6/11754504/20bbe118e30f/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cf6/11754504/8a7cd6ea3e02/gr6.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cf6/11754504/a973aa079ccf/fx1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cf6/11754504/03a95a01d59f/gr1.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cf6/11754504/7d3a124ea6db/gr2.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cf6/11754504/183218e508ce/gr3.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cf6/11754504/1a0f609c0ab9/gr5.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cf6/11754504/20bbe118e30f/gr4.jpg
https://cdn.ncbi.nlm.nih.gov/pmc/blobs/3cf6/11754504/8a7cd6ea3e02/gr6.jpg

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本文引用的文献

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ΔNp63 regulates MDSC survival and metabolism in triple-negative breast cancer.ΔNp63调节三阴性乳腺癌中髓源性抑制细胞的存活和代谢。
iScience. 2024 Feb 29;27(4):109366. doi: 10.1016/j.isci.2024.109366. eCollection 2024 Apr 19.
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Regulation of ROS in myeloid-derived suppressor cells through targeting fatty acid transport protein 2 enhanced anti-PD-L1 tumor immunotherapy.通过靶向脂肪酸转运蛋白 2 调节髓源性抑制细胞中的 ROS 增强抗 PD-L1 肿瘤免疫治疗。
Cell Immunol. 2021 Apr;362:104286. doi: 10.1016/j.cellimm.2021.104286. Epub 2021 Jan 19.
3
The Role of Myeloid-Derived Suppressor Cells (MDSC) in Cancer Progression.
髓源性抑制细胞（MDSC）在癌症进展中的作用
Vaccines (Basel). 2016 Nov 3;4(4):36. doi: 10.3390/vaccines4040036.
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