Abstract
Current knowledge on brain oligodendrogenesis, composed of three sequential waves from different regions, is mainly obtained from rodent studies. Oligodendroglial development and myelination in the rodent brain may not fully mirror those processes in the human brain. Here, we provide a step-by-step protocol for generating fused forebrain organoids derived from human pluripotent stem cells. These fused organoids recapitulate human oligodendroglial developments, offering innovative insights into human myelination research in vitro. For complete details on the use and execution of this protocol, please refer to Cameron-Curry and Le Douarin (1995), Kessaris et al. (2006), and Kim et al. (2019).