As shown in Fig. 4A, after 14 days culture in differentiation medium, organoids became more dense, had thicker outer walls and stopped proliferating as shown in Fig. 4B, at gene expression level, stem cell and proliferation markers LGR5 and Ki76 were significantly downregulated in DM-chol both for ECO and ICO, confirming loss of stemness and proliferation. Here, LGR5 is linked to endocrine-cerebro-osteodysplasia syndrome.