Primary Culture of Adult Neural Progenitors

In this chapter, we describe methodology for in vitro culture of adult neural stem and progenitor cells. The mammalian adult brain, once thought to be completely postmitotic, is now recognized to contain a finite number of neural stem cells, progenitor cells with the capacity for self-renewal and the ability to differentiate into functional neurons and glia (1 ). The largest populations are found in the subventricular zone (SVZ) of the lateral ventricle and the subgranular zone (SGZ) of the dentate gyrus (1 ). It is predicted that these populations, as well as smaller pools of stem and progenitor cells in other brain regions, will be affected by chronic exposure to drugs of abuse. In support of this hypothesis, psychomotor stimulants and opioids have been shown to influence activation of neural progenitors in adult tissue in vitro (2 ) and in vivo (3 ). To xic dopaminergic insult by repeated exposure to 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP) increases progenitor proliferation and subsequent gliogenesis (4 ). Chronic prenatal exposure to drugs of abuse such as cocaine impairs neurogenesis and migration of differentiating neurons (5 ,6 ). One means of investigating the underlying molecular mechanisms responsible for these effects on neural precursors is through in vitro culture. In the protocol provided in the following subheadings, a methodology for the generation of primary stem and progenitor neurospheres from the subventricular zone of the C57Bl/6 mouse is described.