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

Year: 2024

Type: article

Abstract: 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 • Dissection of human embryonic yolk sac and embryo proper • In vitro differentiation assay of human embryonic cells under ILC-induced condition • Steps for conducting CITE-seq using cultured ILCs • Identification of characteristics and functions of various cultured ILC subsets Publisher’s note: Undertaking any experimental protocol requires adherence to local institutional guidelines for laboratory safety and ethics. 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.

Source: STAR Protocols

DOI: 10.1016/j.xpro.2024.103525

Authors: Chen Liu, Qingfeng Fu, Guoju You, Tao Cheng, Yu Lan, Bing Liu