PMC

Nuclei are counter-stained with DAPI. Scale bar, 50 μm.

Immunofluorescence stains of engrafted human GFP spheroids in the small intestine on post-implantation week 1. (a) E-cadherin, (b) lysozyme, (c) Muc2, (d) chromogranin A and (e) CD10. Nuclei are counterstained blue with DAPI. Scale bars, 50 μm.

(A) Cells were implanted into a jejunal segment (hatched area) that was in continuity of the small intestine by isolating a mucosal pocket using a semicircular clamp. (B) Alternatively, cells were injected into a bypassed jejunal segment that did not receive the regular stream of bowel content (hatched area). The arrows show direction of the flow of the ingesta.

(a-b) Brightfield images in culture; (c-j) immunohistochemical analysis, (c-d) lysozyme, (e-f) PAS, (g-h) synaptophysin, (i-j) CD10. The majority of the tested differentiation markers was not visualized in spheroids. Stained structures were obtained from a GFP mouse and showed expected variability in GFP expression. Scale bars, 50 μm.

(a) one week after implantation; (b-f) four weeks after implantation: (b) E-cadherin, (c) lysozyme, (d) synaptophysin, (e) CD10, (f) Muc2. (g-h) H&E stain and GFP immunofluorescence of consecutive slides of engrafted mouse intestine four weeks post-implantation. Nuclei are counter-stained with 4',6-diamidino-2-phenylindole (DAPI). Scale bars, 20 μm (a-f) and 50 μm (g-h).