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Identification of culture conditions for inducing neuroectoderm and mesendoderm differentiation in CDM-PVA.

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posted on 21.02.2013, 05:30 by Ludovic Vallier, Thomas Touboul, Zhenzhi Chng, Minodora Brimpari, Nicholas Hannan, Enrique Millan, Lucy E. Smithers, Matthew Trotter, Peter Rugg-Gunn, Anne Weber, Roger A. Pedersen

H9 hESCs were grown for different periods in CDM-PVA supplemented with different growth factors (Activin, FGF2 or BMP4) or chemical inhibitors SU5402 (SU) for FGF. Then, immunostaining analyses were used to determine the fraction of cells expressing the pluripotency marker Oct-4, the neuroectoderm marker Sox2, the mesoderm marker Brachyury, and the endoderm marker Sox17. The levels of marker-expressing cells were divided into five arbitrary categories: 0 for absence of expression, Very Low (VL) for <1% expressing cells, Low (L) for <5% expressing cells, Moderate (M) for >10% expressing cells, High (H) for >50% expressing cells. Indicated in blue are conditions allowing the generation of neuroectoderm (expressing Sox2) and in red those inducing mesendoderm (cells expressing Brachyury or Sox17).