Assessment of Human Brain Organoids Maturation in Ex-Utero Culture Conditions

Differentiation of human pluripotent stem cells to brain-like structures termed organoids offers an unprecedented opportunity to model human brain development and disease. Brain organoids are self-organized 3D neural tissue that recapitulate some aspects of early brain development. Despite the recent progress in brain organoid technology, current brain organoid protocols and culture conditions generate brain organoids that recapitulate early embryonic stages but are insufficient to promote maturation to postnatal stages. This limitation restricts broader applications in investigating disease mechanisms. Here we aim to address a fundamental limitation of current brain organoids systems by trialing potential culture conditions that could promote advanced maturation of brain organoids. To start addressing this question, we adapted a recently established Roller culture-based platform that supports prolonged ex-utero growth of both natural and mESC-derived synthetic embryos beyond post- gastrulation stages. Our hypothesis is that optimizing embryo-compatible Roller culture conditions may enhance the maturation of brain organoids in vitro.