In vitro expansion of fetal neural progenitors as adherent cell lines.

In vitro studies of neural progenitors isolated from the developing mouse have provided important insights into intrinsic and extrinsic pathways that control their behavior. However, use of primary cultures or neurospheres established from fetal tissues in cell population-based assays can be compromised by cellular heterogeneity. A complementary approach that addresses this issue is the establishment of adherent clonal neural stem (NS) cell lines. Here I describe protocols and troubleshooting advice for establishing adherent NS cell lines from the mouse fetal forebrain. NS cells grow as pure cultures in defined serum-free conditions as adherent monolayers and are therefore amenable to chemical/genetic screens, biochemical studies, and population-based analysis of gene expression or transcriptional regulation (e.g. RNA-Seq and ChIP-Seq). NS cell lines therefore represent a tractable cellular model system to explore the molecular and cellular biology of neural stem cell self-renewal and differentiation. Similar protocols can be extended to rat and human embryos, as well as human brain tumors.