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Our results suggest that such a gradient of signals for differentiation is generated and maintained through cell-cell interactions within the system.
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A list of genes associated with differentiation was generated with MedscanReader 2.2 (Ariadne Genomics) and Pathway Studio 6.2 (Ariadne Genomics) software that enables the automated extraction of information from scientific text (list displayed as supplementary data).
In fact, there are several established experimental results supporting the idea that signals controlling differentiation are generated within the system.
Using various data mining approaches, lists of stem cell-enriched genes and genes that are induced during the in vitro equivalent of epiblast and primitive streak stages of differentiation were generated.
This delayed cell does not receive the stem cell niche signaling anymore (because it is detached from the niche), but it also does not receive the signal to differentiate that, as we propose here, is generated in the differentiation niche (ECs) in response to ecdysone signaling.
Plasma growth differentiation factor-15 (GDF-15) is generated as a 40-KDa propeptide from which the NH2-terminus is cleaved and the resulting 30-KDa protein is secreted as active form [ 138].
These results suggest that typical NPCs with integrated differentiation potentials are generated using this strategy.
This approach had the disadvantage that only immature differentiation stages were generated and the relatively small cell numbers derived in colonies severely limited subsequent investigations.
(3) Effect of zebularine on mESC differentiation: EBs were generated as in experimental point 2 and the cells were treated with 30 μM of zebularine for 7 days (see Figure 2a).
Second, we showed that the observed within-cohort genetic differentiation can be generated by a spatio-temporal subdivision of breeders in distinct reproductive groups constituted by a limited effective number of spawning eels (of the order of few hundred individuals).
For in vitro differentiation, EBs were generated as described above, grown for 8 days in suspension, placed on Matrigel-coated glass coverslips, and allowed to further differentiate for 8 days in mTeSR1 medium.
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