Identification and Maintenance of Neural Precursors from Human Embryonic Stem Cells

This chapter reviews the scientific background of the development of neural precursors from human embryonic stem (hES) cells and describes methods for isolation, propagation, and characterization of hES cell-derived neural precursors. Highly enriched cultures of neural precursors may be derived from the cells. The neural precursors can proliferate in culture for prolonged periods of time, and can differentiate in vitro into mature neurons, astrocytes, and oligodendrocytes. The neurons that are generated respond to neurotransmitter application, their cell membranes contain voltage-dependent channels, and they generate action potentials in response to depolarizing stimuli. The precursors respond to host brain signal, as suggested by their migration along the established host brain migratory tracks and their differentiation in a regional specific manner to progeny of the three neural lineages. The establishment of neuroectodermal precursors from hES cells provides advantages for basic and applied studies of human central nervous system development and disease. Transplantation studies are useful for the analysis of the capability of neural precursors to migrate and differentiate in response to host parenchymal cues and to participate in the processes of host brain development. The chapter also describes the methodology for induction of differentiation in vitro and for the immunophenotyping of differentiated progeny, and describes some of the most commonly used markers.