Neural stem cells, with the capacity of self-renewal and abilities to differentiate into the major cell types of the brain, exist in the developing and adult rodent central nervous system. These cells can be grown in vitro for long periods of time while retaining the capacity to differentiate into neurons, astrocytes, and oligodendrocytes. In this chapter, we describe the steps in detail to isolate and expand neural stem cells in the form of neurospheres from tissue dissections of the embryonic mouse brain. Procedures for the long-term passage of neurospheres, the cryopreservation of neurospheres, and their differentiation are also provided. Although the methodology seems simple, strict adherence to the procedures described is required in order to achieve reliable and consistent results.
Heldmann U, Mine Y, Kokaia Z, Ekdahl CT, Lindvall O (2011) Selective depletion of Mac-1-expressing microglia in rat subventricular zone does not alter neurogenic response early after stroke. Exp Neurol 229:391–398CrossRefPubMedGoogle Scholar
Jagasia R, Song H, Gage FH, Lie DC (2006) New regulators in adult neurogenesis and their potential role for repair. Trends Mol Med 12:400–405CrossRefPubMedGoogle Scholar
Limke TL, Rao MS (2002) Neural stem cells in aging and disease. J Cell Mol Med 6:475–96Google Scholar
Reynolds BA, Weiss S (1992) Generation of neurons and astrocytes from isolated cells of the adult mammalian central nervous system. Science 255:1707–1710CrossRefPubMedGoogle Scholar
Seri B, Garcia-Verdugo JM, McEwen BS, Alvarez-Buylla A (2001) Astrocytes give rise to new neurons in the adult mammalian hippocampus. J Neurosci 21:7153–7160PubMedGoogle Scholar
Weiss S, Reynolds BA, Vescovi AL, Morshead C, Craig CG, van der Kooy D (1996) Is there a neural stem cell in the mammalian forebrain? Trends Neurosci 19:387–393CrossRefPubMedGoogle Scholar