RT Journal Article
T1 Optimized Protocol for Characterization of Mouse Gut Innate Lymphoid Cells
A1 Valle Noguera, Ana
A1 Gómez Sánchez, María José
A1 Girard Madoux, Mathilde J. H.
A1 Cruz Adalia, Aranzazu
AB Since their discovery, innate lymphoid cells (ILCs) have gradually been gaining greater relevance in the field of immunology due to their multiple functions in the innate immune response. They can mainly be found in mucosal and barrier organs like skin, gut, and lungs, and have been classified into five main types (NKs, ILC1s, ILC2s, ILC3s, and Lti cells) according to their function and development. They all play major roles in functions such as tissue homeostasis, early pathogen defense, regulation of inflammation, or tissue remodeling. ILCs are mostly tissue-resident cells tightly bound to the tissue structure, a fact that requires long and complex protocols that do not always provide sufficient yield for analysis. This suggests the need for optimized approaches aimed at ensuring that enriched and viable ILC samples are obtained, in order to furnish quality results. Herein a detailed protocol is established for obtaining a single-cell suspension highly enriched in lymphoid cells from mouse gut in order to identify the different subsets of ILCs by means of flow cytometry. The cell marker panel and flow cytometry gating strategies for identification and quantification of all the different ILC populations are provided for simultaneous analysis. Moreover, the protocol described includes a procedure for studying the different cytokines produced by ILC3s involved in maintaining the integrity of the gut barrier and defending against extracellular pathogens. As a result, herein an efficient method is presented for studying mouse ILCs within the lamina propria of the small intestine and colon; this can constitute a useful tool for future investigations in the field.
PB Frontiers Media
SN 1664-3224
YR 2020
FD 2020-11-30
LK https://hdl.handle.net/20.500.14352/92149
UL https://hdl.handle.net/20.500.14352/92149
LA eng
NO Valle-Noguera A, Gómez-Sánchez MJ, Girard-Madoux MJH, Cruz-Adalia A. Optimized Protocol for Characterization of Mouse Gut Innate Lymphoid Cells. Front Immunol. 2020 Nov 30;11:563414. doi: 10.3389/fimmu.2020.563414. PMID: 33329525; PMCID: PMC7735015
NO Ministerio de Ciencia, Innovación e Universidades
NO Agenda Estatal de Investigación
NO Fondo Europeo de Desarrollo Regional
DS Docta Complutense
RD 17 jul 2025