Protocol for in vitro generation of innate lymphoid cells from human embryonic tissues

Deciphering the origins of innate lymphoid cells (ILCs) is critical for a deeper understanding of innate immunity. Here, we present a protocol for assessing ILC potential of human embryonic resources. We describe steps for identifying lymphoid progenitors in human embryonic tissues, then culturing mature ILCs in vitro, and identifying characteristics and functions of different cultured ILC subsets using cellular indexing of transcriptomes and epitopes (CITE)-sequencing and flow cytometry analysis. This protocol provides a simple platform for investigating the development of human embryonic ILCs. For complete details on the use and execution of this protocol, please refer to Ni et al.¹.