Isolating and culturing of precursor cells from the adult human brain

Adult neural precursor cells are an essential part of the brain, and a focus of two decades of intense research (Ming and Song, Neuron 70:687-702, 2011). Even though adult human stem/progenitor cells have been identified early on (Kirschenbaum et al., Cereb Cortex 4:576-589, 1994; Eriksson et al., Nat Med 4:1313-1317, 1998), progress in the field of adult human neurogenesis has been slow. The reasons for this may be more advanced neighboring fields of pluripotent stem cell research, and lacking study material as well as well-established and standardized protocols. Furthermore, adult precursor cells in humans seem to have greater potential than in rodents (Walton et al., Development 133:3671-3681, 2006). This may be attributed to species differences in astrocyte development and diversity (Oberheim et al., Neurosci 29:3276-3287, 2009). In this chapter, we provide a guideline for adult human brain tissue dissociation, be it from biopsy or autopsy specimens. This is by no means the only way of culturing adult neural precursor cells, but it may help in streamlining research on this fascinating topic, as well as help introducing others into this field. We describe our methodology for establishing and maintaining long-term cultures from white and grey matter, as well as a simple protocol for differentiating these cells.