Generating CNS Neurons from Embryonic, Fetal, and Adult Stem Cells

Abstract

The in vitro culturing of neuroepithelial stem cells has become an indispensable tool for studying the mechanisms controlling proliferation, mitotic arrest, and lineage commitment of cells in the nervous system. However, any potential clinical use of these cells requires systematic methods to enrich for the cell of interest and demonstrate that these cells show functions that will assist in understanding and treating the disease. Embryonic stem (ES) cells are pluripotent and immortal cells derived from the inner cell mass of preimplantation blastocysts. They can proliferate extensively and have the ability to differentiate into endodermal, mesodermal, and ectodermal derivatives. The most important benefit of using ES cells as donor cells is the relative ease of genetic engineering, which permits the enrichment or purification for specific cell types by selectable marker. The efficient methods of generating central nervous system (CNS) stem cells and their derivatives from ES cells have been developed. Primary fetal and adult CNS stem cells are also extensively self-renewing and are multipotent for neuronal and glial fates. This chapter discusses the methods of culturing primary CNS stem cells, as they have formed the basis for all subsequent methods of generating and comparing CNS derivatives from ES cells.