Isolation of adoptively transferred CD8+ T cells in mouse tumor tissues for lipid peroxidation detection

Liuling Xiao; Qing Yi

doi:10.1016/j.xpro.2022.101945

Isolation of adoptively transferred CD8⁺ T cells in mouse tumor tissues for lipid peroxidation detection

Liuling Xiao, Qing Yi

Research output: Contribution to journal › Article › peer-review

3 Scopus citations

Abstract

The lipid peroxidation level of tumor-infiltrating CD8⁺ T cells is crucial for its activity and longevity. Here, we describe a protocol for effective and epitope-preserving dissociation of mouse tumors and subsequent leukocyte purification and lipid peroxidation staining of adoptively transferred CD8⁺ T cells. We use BODIPY 581/591 C11 to monitor the cellular lipid peroxidation level and detect its fluorescent change by flow cytometry, followed by analysis in FlowJo. This protocol is adaptable to intrinsic CD8⁺ T cells in tumors as well. For complete details on the use and execution of this protocol, please refer to Xiao et al. (2022)¹ and Ma et al. (2021).²

Original language	English (US)
Article number	101945
Journal	STAR Protocols
Volume	4
Issue number	1
Early online date	Dec 15 2022
DOIs	https://doi.org/10.1016/j.xpro.2022.101945
State	Published - Mar 17 2023

Keywords

Cancer
Cell Biology
Flow Cytometry/Mass Cytometry
Immunology
Metabolism
Molecular/Chemical Probes
Single Cell

ASJC Scopus subject areas

General Neuroscience
General Biochemistry, Genetics and Molecular Biology
General Immunology and Microbiology

Access to Document

10.1016/j.xpro.2022.101945

Cite this

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title = "Isolation of adoptively transferred CD8+ T cells in mouse tumor tissues for lipid peroxidation detection",

abstract = "The lipid peroxidation level of tumor-infiltrating CD8+ T cells is crucial for its activity and longevity. Here, we describe a protocol for effective and epitope-preserving dissociation of mouse tumors and subsequent leukocyte purification and lipid peroxidation staining of adoptively transferred CD8+ T cells. We use BODIPY 581/591 C11 to monitor the cellular lipid peroxidation level and detect its fluorescent change by flow cytometry, followed by analysis in FlowJo. This protocol is adaptable to intrinsic CD8+ T cells in tumors as well. For complete details on the use and execution of this protocol, please refer to Xiao et al. (2022)1 and Ma et al. (2021).2",

keywords = "Cancer, Cell Biology, Flow Cytometry/Mass Cytometry, Immunology, Metabolism, Molecular/Chemical Probes, Single Cell",

author = "Liuling Xiao and Qing Yi",

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