The infiltration of activated T cells, such as CD8+ effector, in metabolic tissues plays a crucial role for the initiation and propagation of obesity-induced inflammation. Given the pivotal role of lactate transporter monocarboxylate transporter 1 (MCT1) in immune cell activation, we present a protocol for the isolation and activation of CD8+ T lymphocytes selectively lacking MCT1. We describe steps for the induction of adipocyte differentiation, CD8+ T isolation and activation, and adipocyte-CD8+ T cell co-culture. We then detail qPCR analysis on differentiated adipocytes. For complete details on the use and execution of this protocol, please refer to Macchi et al.1.

Protocol to evaluate the impact of murine MCT1-deficient CD8+ T cells on adipogenesis / C. Macchi, A. Moregola, G.D. Norata, M. Ruscica. - In: STAR PROTOCOLS. - ISSN 2666-1667. - 4:2(2023 Jun 16), pp. 102301.1-102301.12. [10.1016/j.xpro.2023.102301]

Protocol to evaluate the impact of murine MCT1-deficient CD8+ T cells on adipogenesis

C. Macchi
Primo
;
A. Moregola;G.D. Norata
Penultimo
;
M. Ruscica
Co-ultimo
2023

Abstract

The infiltration of activated T cells, such as CD8+ effector, in metabolic tissues plays a crucial role for the initiation and propagation of obesity-induced inflammation. Given the pivotal role of lactate transporter monocarboxylate transporter 1 (MCT1) in immune cell activation, we present a protocol for the isolation and activation of CD8+ T lymphocytes selectively lacking MCT1. We describe steps for the induction of adipocyte differentiation, CD8+ T isolation and activation, and adipocyte-CD8+ T cell co-culture. We then detail qPCR analysis on differentiated adipocytes. For complete details on the use and execution of this protocol, please refer to Macchi et al.1.
Cell Biology; Cell culture; Cell isolation; Flow Cytometry/Mass Cytometry; Metabolism; Model Organisms; Molecular Biology
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16-giu-2023
19-mag-2023
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Utilizza questo identificativo per citare o creare un link a questo documento: https://hdl.handle.net/2434/979269
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