Generation of Schwann cells and sensory neurons from human embryonic stem cells

This protocol describes a method to generate peripheral sensory neuron-like cells and Schwann-like cells by co-culturing human embryonic stem cells with PA6 stromal cells. In the first step, neurospheres expressing neural crest markers are produced. When plated on PA6 cells and cultured in neural induction medium, adherent neurospheres differentiate into peripheral neuron-like cells. After 1 week of culturing on PA6 feeder cells, 25% of the cells express peripheral sensory neuron (PSN)-like markers. Prolonged culturing on PA6 feeder cells leads to expression of PSN markers, voltage-dependent current activity and action potentials. Alternatively, neurospheres can be cultured on poly-lysine/laminin-coated coverslips as aggregates in a specific differentiation medium, to form Schwann-like cells. These cells express unique Schwann cell-markers, produce myelin and wrap axons in in-vitro co-cultures.