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

Figure 1 Characterization of the pluripotency and haploidy of extended pluripotent human haploid embryonic stem cells. A: The human diploid embryonic stem cell (ESC) line H9 (left) was used as a control for extended human haploid ESCs (haESC) (right). Scale bar, 100 μm; B: Fluorescence activated cell sorting analysis of the DNA content of the human haESC line hPGES1 after approximately 6 wk of culture, without sorting. Left, diploid ESC line as a control; Right, haploid cell line EhPGES1. The “n” peak represents haploid cells; C: G-band analysis of the EhPGES1 cell line with a haploid set of 23 chromosomes (right) and diploid human ESCs as a control (46 chromosomes, left); D: Alkaline phosphatase staining of the human haESC line EhPGES1 (right) and diploid human ESCs as a control (left). Scale bar, 200 μm; E: Immunofluorescence analysis of the primate ESC markers Oct4 (red), Nanog (green) and SSEA4 (green) in the human haESC line EhPGES1. DAPI (blue) was used to stain the nuclei. Scale bar, 100 μm; F: Morphology of embryoid bodies from haploid-enriched cells (right) and diploid human ESCs as a control (left). Scale bar, 100 μm.