Human fetal brain self-organizes into long-term expanding organoids

Human brain development involves an orchestrated, massive neural progenitor expansion while a multi-cellular tissue architecture is established. Continuously expanding organoids can be grown directly from multiple somatic tissues, yet to date, brain organoids can solely be established from pluripotent stem cells. Here, we show that healthy human fetal brain in vitro self-organizes into organoids (FeBOs), phenocopying aspects of in vivo cellular heterogeneity and complex organization. FeBOs can be expanded over long time periods. FeBO growth requires maintenance of tissue integrity, which ensures production of a tissue-like extracellular matrix (ECM) niche, ultimately endowing FeBO expansion. FeBO lines derived from different areas of the central nervous system (CNS), including dorsal and ventral forebrain, preserve their regional identity and allow to probe aspects of positional identity. Using CRISPR-Cas9, we showcase the generation of syngeneic mutant FeBO lines for the study of brain cancer. Taken together, FeBOs constitute a complementary CNS organoid platform. [Display omitted] •Human fetal brain organoids (FeBOs) display cellular heterogeneity and can be expanded•FeBOs produce a tissue-like ECM niche and enable ECM perturbation studies•Derivation of regional FeBOs allows the study of regional morphogen effects•CRISPR-engineered FeBOs are a scalable bottom-up tumor modeling platform Tissue-derived human fetal brain organoids (FeBOs) are established from different anatomical regions that can be expanded in culture. FeBOs may help understand developmental and disease-related biology.