Figure 1

Human central nervous system stem cell (HuCNS-SC) differentiation in vitro. Human cells plated in culture become specific neuronal subtypes under defined growth factor conditions. (A) Gabaergic (gamma-aminobutyric acid, GABA); (B) dopaminergic (tyrosine hydroxylase; TH); (C) cholinergic (choline acetyltransferase; ChAT). (D) Differentiated neurons mature in vitro as shown by voltage-activated sodium and potassium currents from a clamp recording. Adapted from [29]. (E,F) HuCNS-SC also differentiate into astrocytes as defined by glial fibrillary acidic protein (GFAP; E) or oligodendrocytes identified by the marker O4 (F). In culture, rare oligodendrocytes can mature to myelin basic protein (MBP)-positive cells (inset of F). (G) Images of fluorescent bead lawn in which tracks cleared of beads by migrating/phagocytosing HuCNS-SC appear as black and beads ingested by the cells appear as bright spots (upper panel), and composite images of beads in blue and phalloidin stained cells in red (lower panel). Cells located within cleared track areas (red in lower panel) co-localize with bright spots of phagocytosed beads in upper panel. (H) Quantification of migration area (fluorescent beadfree) with high-content assay analysis software. Factors in fetal bovine serum (FBS) significantly enhanced migratory/phagocytosing activity, while it was blocked by the actin polymerisation inhibiting reagent cytochalasin D (CytD). Data represents the results of three independent experiments in triplicate wells.