Primary culture and live imaging of adult neural stem cells and their progeny.

Adult neural stem cells (NSC) generate neurons throughout life, but little is known about the sequence of events involved in the transition from NSC to neurons. Studying the intermediary steps involved in the specification of neuronal cells from NSCs requires observation of cells in real time. Here we describe a primary culture of the adult subependymal zone (SEZ) which allows for continuous live imaging to characterize the mode of cell division and lineage progression of adult NSCs and their progeny. To this end, cells are cultured at low density under adherent conditions and without growth factors. Under these conditions, NSCs display classical hallmarks of adult SEZ NSCs in vivo, such as astroglial marker expression and promoter activity, a slow cell cycle, and a predominantly neurogenic potential. Video time-lapse microscopy experiments using this cell preparation allow for studying the steps involved in the generation of fast-dividing precursors and neuroblasts from slow-dividing astroglia/NSCs.