Generation of a human haploid neural stem cell line for genome-wide genetic screening

Figure 2 Derivation of human haploid neural stem cells via differentiation invitro. A: Schematic overview of the strategy to efficiently derive haploid neural stem cell (haNSC) lines through embryoid body-mediated differentiation and fluorescence activated cell sorting; B: Morphology of 7-d aggregates. Scale bar, 100 μm; C: Morphology of primitive neuroepithelia. Dashed lines indicate the morphology of primitive neuroepithelia. Scale bar, 100 μm; D: Morphology of definitive neuroepithelia. Dashed lines indicate the morphology of definitive neuroepithelia. Scale bar, 100 μm; E: Fluorescence activated cell sorting of passage 4 human haNSCs and derived haNSCs maintained a haploid DNA content over 10 passages. Left, a diploid cell line as a control; middle, passage 4 haNSCs; right, derived haNSCs after 10; F: Morphology of sorted haNSCs. Scale bar, 200 μm; G: Morphology of haploid neural spheres from sorted haNSCs. Scale bar, 200 μm; H: G-band analysis of haNSCs with a haploid set of 23 chromosomes. haNSCs: Haploid neural stem cell; haESCs: Haploid embryonic stem cells; FACS: Fluorescence activated cell sorting; EB: Embryoid body; NPC: Neural progenitor cells.