Supplementary Figure 3: Isolation and ex vivo expansion of mouse BM-derived ILC2 cells. | Nature Immunology

Supplementary Figure 3: Isolation and ex vivo expansion of mouse BM-derived ILC2 cells.

From: Type I interferon restricts type 2 immunopathology through the regulation of group 2 innate lymphoid cells

Supplementary Figure 3

(a) Schematic representation of isolation and ex vivo expansion of murine bone marrow-derived group 2 innate lymphoid cells. (b) Bone marrow cells were stained for Sca-1, CD117 (c-Kit), CD25, and for lineage markers (TCRβ, TCRγδ, CD3ɛ, Gr-1, CD11b, TER-119, B220, NK1.1, CD5, CD11c) and lineage-negative Sca-1+c-Kit-CD25+ group 2 innate lymphoid cells were enriched by flow cytometric cell sorting. (c) Purity and (d) expression of CD127, T1/ST2, ICOS, KLRG1, CD90.2 and Gata3 of sorted bone marrow-derived ILC2 was analyzed by flow cytometry. Cells were then cultured in the presence of IL-2, IL-7, IL-25, IL-33 (all at 50 ng/ml), and TSLP (20 ng/ml) for 15 days. Cells were given fresh medium and cytokines every two days and split into multiple wells when necessary to avoid excessive crowding, yielding an increase in ILC2 numbers of approximately 500-fold. The resulting cells were then seeded in fresh medium and allowed to rest in IL-2 and IL-7 (both at 10 ng/ml) for two days. Medium was removed from wells and rested cells were stimulated as necessary for up to five days.

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