hpESCs generate NSCs.

<p>(<b>A</b>) Time-lapse, phase-contrast images illustrating the individual stages of neural <i>in vitro</i> differentiation of hpESC line LLC9P towards hpNSCs. Starting from hpESCs grown on human foreskin fibroblasts to hpESC-derived floating embryoid bodies, attached embryoid bodies (insert indicates rosette-like structures), floating neurospheres and NSCs. Scale bars, left panel: 0.5 mm; other panels: 0.25 mm. (<b>B</b>) RT-PCR analysis for the expression of pluripotency (<i>Oct4</i> and <i>Nanog</i>) and neural stem cell markers (<i>Sox1</i>, <i>Nestin</i>, <i>Pax6</i> and <i>MS1</i>) in undifferentiated hESC and hpESC cultures, and in hNSC and hpNSC cultures. Also shown is a RT-PCR analysis for the expression of the neural crest cell markers <i>Snai2</i> and <i>FoxD3</i> and the mesodermal marker <i>Acta1</i> in undifferentiated hESCs, hNSCs, hpESCs, and hpNSCs at passages 5, 10 and 15. Controls shown are analyses of human parthenogenetic neural crest stem cells (hpNCSCs), human fetal brain (hFB) and human adipose tissue-derived mesenchymal stromal cells (hMSCs). (<b>C</b>) Representative confocal images of hpNSC cultures immunostained with antibodies specific for <i>Nestin</i>, <i>Sox1</i>, <i>Sox2</i>, and <i>Vimentin</i> are shown. Scale bars: 50 µm; n = 3.