To date, several approaches have been developed for differentiating pluripotent stem cells into cortical neurons. Some of these methods achieve neural induction in high-density monolayer cultures10 or by embedding clusters of hiPSCs in gelatinous protein mixtures (such as Matrigel) and later culturing them in a spinning bioreactor8. Other approaches use embryoid bodies derived from hiPSCs that are either plated on coated surfaces to generate neural progenitors organized in rosettes32–34 or maintained in suspension initially in serum-free conditions and later in serum and Matrigel (for example, SFEBq: serum-free floating culture of embryoid body–like aggregates with quick reaggregation)6,7. Here we describe an approach for in vitro neural differentiation of human pluripotent stem cells that has several advantages.
