The method is simple and does not involve cell (re)-plating, embedding into extracellular matrices or culture in complex environments. hCSs are maintained in floating conditions on low-attachment plates with biweekly changes of regular serum-free Neurobasal with B-27 medium (Invitrogen), and can be easily maintained for up to 9 months in vitro. Moreover, the specification of hCS, as well as the functional properties of the resulting cells, are reproducible between hiPSC lines and differentiation experiments.
