Objective To investigate the isolation and culture of the type 2 innate lymphoid cell (ILC2) in the lungs of newborn mice. Methods Immunomagnetic bead enrichment and fluorescence activated cell sorting (FACS) were used to isolate ILC2s. Flow cytometry was used to identify the purity of ILC2s. Inverted microscope was used to observe the growth of cells. ELISA was used to detect the expression levels of interleukin 5 (IL-5) and IL-13. Results The purity of the isolated ILC2s reached more than 95%. The isolated ILC2s were round or oval, and suspended in cell culture medium. After stimulation with IL-2 and IL-7 combined with IL-33, the contents of IL-5 and IL-13 produced by ILC2s and the proliferation ability of ILC2s increased significantly. Conclusion A rapid and efficient method for isolation and culture of ILC2s in the lung of newborn mice has been found.