Protocol for in vitro isolation, induction, expansion, and determination of human natural regulatory T cells and induced regulatory T cells

Jian Gu; Qing Shao; Jinren Zhou; Qiuyang Chen; Ling Lu

doi:10.1016/j.xpro.2022.101740

Protocol for in vitro isolation, induction, expansion, and determination of human natural regulatory T cells and induced regulatory T cells

STAR Protoc. 2022 Dec 16;3(4):101740. doi: 10.1016/j.xpro.2022.101740. Epub 2022 Oct 9.

Authors

Jian Gu¹, Qing Shao¹, Jinren Zhou¹, Qiuyang Chen¹, Ling Lu²

Affiliations

¹ Hepatobiliary Center of the First Affiliated Hospital, Nanjing Medical University, Research Unit of Liver Transplantation and Transplant Immunology, Chinese Academy of Medical Sciences, Nanjing, Jiangsu 210029, China.
² Hepatobiliary Center of the First Affiliated Hospital, Nanjing Medical University, Research Unit of Liver Transplantation and Transplant Immunology, Chinese Academy of Medical Sciences, Nanjing, Jiangsu 210029, China. Electronic address: lvling@njmu.edu.cn.

Abstract

Regulatory T cells (Tregs) can inhibit the occurrence of autoimmune diseases and increase the activation threshold of the immune response. Here, we present schemes for the isolation and culture of natural human regulatory T cells (nTregs) and in vitro-induced Tregs (iTregs). Appropriate concentrations of TGF-β, IL-2, retinoic acid (atRA), and rapamycin were used to promote proliferation to meet sample needs in basic research, especially in technologies such as sequencing. For complete details on the use and execution of this protocol, please refer to Lu et al. (2014a) and Gu et al. (2022).

Keywords: Cell biology; Cell culture; Cell isolation; Immunology.

Publication types

Research Support, Non-U.S. Gov't

MeSH terms

Humans
Sirolimus
T-Lymphocytes, Regulatory*
Transforming Growth Factor beta*

Substances

Transforming Growth Factor beta
Sirolimus