Intracellular ATP delivery to in vitro expanded mouse CD27- γδ T cells

Yunke Wang; Jun Yan

doi:10.1016/j.xpro.2023.102532

Intracellular ATP delivery to in vitro expanded mouse CD27^- γδ T cells

STAR Protoc. 2023 Sep 15;4(3):102532. doi: 10.1016/j.xpro.2023.102532. Epub 2023 Aug 25.

Authors

Yunke Wang¹, Jun Yan²

Affiliations

¹ Division of Immunotherapy, The Hiram C. Polk, Jr., MD Department of Surgery, Immuno-Oncology Program, Brown Cancer Center, University of Louisville School of Medicine, Louisville, KY 40202, USA; Department of Head and Neck Radiotherapy, The Cancer Hospital of the University of Chinese Academy of Sciences (Zhejiang Cancer Hospital), Zhejiang 310022, China.
² Division of Immunotherapy, The Hiram C. Polk, Jr., MD Department of Surgery, Immuno-Oncology Program, Brown Cancer Center, University of Louisville School of Medicine, Louisville, KY 40202, USA. Electronic address: [email protected].

Abstract

Intracellular ATP supports the function of γδT17 cells in mice. Here, we present a protocol for intracellular ATP delivery to in vitro expanded mouse CD27^- γδ T cells. We describe steps for pre-coating well plates, preparing lymphocytes, culturing CD27^- γδ T cells, and ATP delivery. We then detail functional evaluation of γδ T cells by flow cytometry. Appropriate concentrations of control and ATP vesicles are detailed for intracellular ATP delivery, which can also be applied to other immune cells. For complete details on the use and execution of this protocol, please refer to Wang et al. (2023).¹.

Keywords: Cell Culture; Cell Isolation; Flow Cytometry/Mass Cytometry; Health Sciences; Immunology; Metabolism.

MeSH terms

Adenosine Triphosphate
Animals
Cells, Cultured
Mice
Receptors, Antigen, T-Cell, gamma-delta*
T-Lymphocytes*

Substances

Receptors, Antigen, T-Cell, gamma-delta
Adenosine Triphosphate