Figure 3 : A. Immunostained HIO derived from F1089A excised spheroids by Protocol I and II excised
spheroids (top and middle panel) and protocol II floating spheroids (bottom panel).

Only Protocol II excised stage 2 spheroids develop into HIO that express moderate- intense immunostaining for E-cadherin,
CDX2 and villin after 36 days of Stage 3 differentiation. B. Stage 1 definitive endoderm marker expression is required for later
stage morphology. Example of an iPSC line with poor definitive endoderm marker expression at Stage 1 that did not develop
excisable spheroids by Stage 2. 4X magnification. C. Enteroendoendocrine marker chromogranin A expression in human
tissue (top) and HIO from A1145A differentiation (bottom).

Hornby et al.Stem Cell Biology and Research  2015 2:1DOI : 10.7243/2054-717X-2-1