FIG 9.

Characterization of bovine ileal enteroids. (A) Representative images of sequential bovine ileal enteroids from intestinal crypts that were grown for 1, 3, or 5 days in complete medium with growth factors. (B) After 10 days of growth in complete medium with growth factors, enteroids typically had two major morphologies: cystic (left) and multilobular (right). (C and D) Differentiated bovine ileal enteroids (3D) grown in Matrigel with a differentiation medium lacking Wnt3A, SB202190, and nicotinamide and with half the normal concentration of Noggin and R-spondin, were fixed with 4% paraformaldehyde and embedded in paraffin. (C) Thin sections were stained with H&E (upper panel), and periodic acid-Schiff (PAS, lower panel). (D) Thin sections were stained using immunohistochemistry to determine the expression levels of villin for enterocytes, chromogranin A for enteroendocrine cells, and mucin 2 for goblet cells. Nuclei were stained with DAPI.