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First, in vitro differentiation protocols are intended to mimic the conditions in the developing embryo and most knowledge on embryonic development of the pancreas was accumulated from mouse studies. Furthermore, the timeline for the specification and maturation of a particular cell is theoretically shorter in mice than humans (respectively 3?weeks and 40?weeks). Second, mESCs are easier to maintain in culture as they grow faster, are more resistant to enzymatic dissociation during passaging, and have less tendency to spontaneously differentiate. mESCs can thus be used as a model to rapidly tweak protocols that thereafter could be implemented on hESCs, for example to obtain beta-cells. However, despite the similarities in their general properties, embryonic stem cells from mouse and human differ in their manipulation in vitro, [http://www.selleckchem.com/products/epz-5676.html EPZ5676] which is related to the developmental origin of these cells. Indeed, in contrast to mESCs, pluripotent cells derived from mouse epiblast stage embryos (epiblast stem cells, EpiSCs) can be considered as the true developmental counterparts of hESCs. Interestingly, both EpiSCs and hESCs were shown to require the same culture conditions (Brons et al., 2007?and?Tesar et al., 2007). Human and mouse ESCs also differ in the conditions needed for definitive endoderm (DE) induction, the first step towards commitment into pancreatic and other gastrointestinal fates (Mfopou et al., 2010b). Whereas hESCs cultured as monolayers and stimulated with Activin A and Wnt3a in a basic medium efficiently generate DE progenitors, mESCs cultured under similar conditions usually fail to survive or they generate DE cells with low efficiency [http://en.wikipedia.org/wiki/MAPK MAPK] ([http://www.selleckchem.com/products/ly2109761.html LY2109761] constitute an optimal environment for efficient differentiation into pancreatic phenotypes (Mfopou et al., 2005?and?Mfopou et al., 2007). Interestingly, monolayer cultures are also technically more practical and simple to rapidly analyze microscopically; e.g. they don't need to be embedded and sectioned. They are thus optimal for high throughput screening of growth factor and small molecule combinations. DE cells were recently generated from mouse embryoid bodies using a combination of ActA (TGF�� activator), Noggin (BMP antagonist), and lithium chloride (Wnt pathway activator) (Li et al., 2011). Whereas ActA and Wnt activators are commonly used for DE induction, Noggin supplementation is justified by the requirement for low BMP signaling to direct the mesendoderm towards anterior primitive streak derivatives (D'Amour et al., 2005, Sumi et al., 2008?and?Wang et al., 2012).