First, experimental optimization showed that HCC-Gelatin and non-adhesive HCC-hydrogel (PEG based)9 structures impose a constraint on growing cystic mSIOs inducing budding and, from day 2 to day 7 of culture, mSIOs LGR5 cell segregation (Supplementary Fig. 4a-c). Here, LGR5 is linked to hepatocellular carcinoma.