Protocol for in vitro generating innate lymphoid cells from mouse α4β7+ lymphoid progenitors

Building a simple and efficient in vitro differentiation system is crucial for studying the regulatory mechanisms during the development of innate lymphoid cells (ILCs). Here, we present a protocol for generating ILC subsets from α₄β₇⁺ lymphoid progenitors (αLPs). We describe steps for murine cell isolation from fetal liver and adult bone marrow, flow cytometry sorting for αLPs, and cell culture. We then detail procedures for flow cytometry analysis of ILCs. This protocol significantly simplifies the differentiation process through ILC differentiation in vitro. For complete details on the use and execution of this protocol, please refer to Wu et al.¹.