Extended Data Fig. 3: Characterization of SiAFO. | Nature Medicine

Extended Data Fig. 3: Characterization of SiAFO.

From: Single-cell guided prenatal derivation of primary fetal epithelial organoids from human amniotic and tracheal fluids

Extended Data Fig. 3

(a) Phase-contrast images showing SiAFOs at passage 6 and their expansion after cryopreservation until passage 10 (scale bar: 200 μm). (b) Whole-mount immunofluorescent staining showing the absence of NKX2-1 (lung) and PAX8 (kidney) in SiAFO (scale bar: 50 μm). (c) Whole-mount immunofluorescent staining for Chromogranin A (CHGA) and Mucin 2 (MUC2) on SiAFOs in expansion medium (scale bar: 50 μm). (d) RT–qPCR analysis of SiAFOs cultured in maturation medium without (Basal), with CHIR99021 (CHIR) or with DAPT (n=5 SiAFO clonal lines from n=2 patients; mean±SEM). (e) Stereoscopic image of an intestinal ring (scale bar: 1 mm) and microCT images showing the presence of budding (left) as well as luminal-like structures (L) (scale bars: 100 μm). (f) 3D z-stacks of SiAFO rings showing tight junctions (ZO-1), Paneth cells (LYZ), enterocytes (FABP1 and KRT20), intestinal secretory cells (MUC2) and enteroendocrine cells (CHGA); nuclei counterstained with Hoechst (scale bars:50μm).

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