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The clonally derived NS cell line NS-5 generated from mouse ES cells was propagated and passaged according to Conti et al. [35] (Fig. 1A).
A similar pattern is apparent in mouse NS cell lines.
These colonies were morphologically undifferentiated and indistinguishable from the parental human NS cell cultures.
The mechanisms and regulation of NS cell dormancy are also of interest for future study.
Colonies could be generated from single cells from all four NS cell populations.
The derivation of human and mouse NS cell lines is described in [26] and [27].
These markers are expressed relatively homogeneously throughout the NS cell population.
Therefore we conclude that Mbd2, Mecp2 and Kaiso are dispensable for NS cell derivation and maintenance.
Since human NS cell cultures do not express differentiation markers under expansion conditions [26], the longer doubling time is unlikely to be due to NS cell differentiation into non-proliferative cell lineages.
We could see no genotype-dependent differences in the efficiency or rate of establishment of NS cell lines.
These changes in mRNA levels would be predicted to have substantial effects on NS cell self-renewal and differentiation.
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