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

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
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

Neuroscience(all)
Biochemistry, Genetics and Molecular Biology(all)
Immunology and Microbiology(all)

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",

note = "Funding Information: This work was funded by National Cancer Institute (NCI) grants R01 CA200539 and R01 CA239255 and the Cancer Prevention & Research Institute of Texas Recruitment of Established Investigator Award ( RR180044 ). Publisher Copyright: {\textcopyright} 2022",

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AU - Yi, Qing

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PY - 2023/3/17

Y1 - 2023/3/17

N2 - 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

AB - 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

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