Fig. 1
Bright-field and H&E images of ISC, ENT, GOB, and PAN organoids. a Images at 20× (top and middle) and 60× (bottom row). There were no differences in morphology of cells in two sets of organoids from different mice. Arrows depict secretory vesicles in GOBs and PANs. b Periodic acid-Schiff/Alcian blue stained images at 20× (top) and 60× (bottom). c Expression profile of biomarkers in TYP, ISC, ENT, GOB, PAN, intestinal crypts (CRY) and tissue homogenates (HOM). TYPs are undirected organoids with all cell types represented. The expression of each biomarker was normalized to that in TYP organoids (=1.0). i The biomarker for stem cells, leucine-rich-repeat-containing G-protein coupled receptor 5 (Lgr5), was highly expressed in ISC organoids (n = 4 for TYP and n = 3 for other organoids). ii PANs are marked by lysozyme (Lyz), which was very abundant in PAN organoids (see scale along ordinate axis) (n = 4 for TYP; n = 3 for others). iii Mucin-2 (Muc2) was highly expressed in GOBs (n = 4 for TYP and n = 3 for others). iv The enteroendocrine biomarker chromogranin A (Chga) was expressed in GOBs and PANs (n = 4 for TYP and n = 3 for others). v ENTs were marked by sucrase isomaltase (SI) (n = 4 for TYP, ISC, and HOM, and n = 3 for others). vi Stem cells were marked by olfactomedin-4 (Olfm4) (n = 4 for TYP, ENT; n = 3 for others). Pa,b,c,d ≤ 0.05. d Immunofluorescence of biomarkers of ISCs, ENTs, GOBs, and PANs. Nuclei are blue. Organoids were stained with OLFM4 (red), lysozyme (LYZ; green), mucin-2 (MUC2; green), sucrase isomaltase (SI; red), or chromogranin A (CHGA; green). 60× magnification; bar = 25 μm. Supporting data sets are deposited in the figshare repository [50]. CHGA chromogranin A, CRY crypt, ENT enterocyte, GOB goblet cell, H&E hematoxylin and eosin stain, HOM homogenates of intestinal mucosa, ISC intestinal stem cell, L lumen, Lgr5 leucine-rich repeat containing G-protein-coupled receptor 5, LYZ lysozyme, MUC2 Mucin-2, OLFM4 Olfactomedin-4, PAN Paneth cell, SI sucrase isomaltase, TYP typical