Abstract
Group 2 innate lymphoid cells (ILC2) are a recently characterized cell population which lacks specific antigen receptors and contributes to immune responses at mucosal surfaces of lung and gut. Recently, we demonstrated that ILC2 expand in the context of chronic liver diseases in mice and contribute to pathology in an IL-33 dependent manner. Here, we describe a protocol to isolate highly purified ILC2 from mouse livers. This procedure provides effective digestion of liver tissue and limits proteolytic degradation of cell surface receptors leading to increased yields of biologically functional cells. The number of liver resident ILC2 in the steady state is low, however their number dramatically increase upon systemic or local treatment of mice with cytokines such as IL-25 and IL-33. Using minicircle-based expression constructs for these cytokines high numbers of functional ILC2 can be isolated with the protocol provided here.
Keywords: Group 2 innate lymphoid cells, ILC2, Inflammation
Materials and Reagents
Equipment
Procedure
Recipes
Note: All materials and reagents should be low endotoxin cell culture quality.
Acknowledgments
This work was supported by the Collaborative Research Center 796 and the Priority program SPP1656 of the DFG (to S.W.) and the Interdisciplinary Center for Clinical Research (IZKF) of the University Medical Center Erlangen.
References
If you have any questions/comments about this protocol, you are highly recommended to post here. We will invite the authors of this protocol as well as some of its users to address your questions/comments. To make it easier for them to help you, you are encouraged to post your data including images for the troubleshooting.