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The persisting reductions of this mark observed 10 days after an acute in vivo exposure, but not within in vitro neurosphere cultures maintained in the stem cell state, suggest recovery of this mark may be dependent upon cells being in a stem cell versus a differentiating state.
To this end, we initiated a treatment protocol wherein cells were maintained in the stem cell state and exposed to varying concentrations of EtOH for 3 days; then allowed to progress through a 4-day recovery period representing at least three population doublings.
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P63 expression has been reported to identify epithelial stem cells [84] however, the abundance of p63-expressing cells in the thymus (Figure 8 & [79], [85], [86], [87], [88]) and other tissues like bladder epithelium indicates that p63 expression is maintained in the immediate progeny of the stem cells as well.
How this quiescence is initiated and maintained in the midst of proliferating stem cells is unknown.
To establish whether pluripotency was maintained in the targeted clones, we evaluated the stem cell marker profile and in vitro differentiation capacity of F344 and SD targeted clones (Figure 2 and Figure S3).
How are stem cells maintained in the right numbers?
Rx2 is expressed in retinal stem cells/RPCs (A ) and after NR differentiation is additionally expressed in photoreceptors and Müller Glia (B ) while its expression is maintained in retinal stem cells of the CMZ (Reinhardt and Centanin et al., submitted).
The cells were maintained in stem cell restricted medium RH-B (Stem Cell Science) containing 2% FCS, with additional insulin (5 μg/ml, Pepro Tech), EGF (10 ng/ml, Pepro Tech) and FGF2 (20 ng/ml, Pepro Tech) for 2 days.
ALDH+ cells were also expanded and maintained in stem cell expansion and undifferentiation media.
SO selective effect on GBM CSCs is corroborated by the lower effectiveness of SO on GBM differentiated cells compared with the same cultures maintained in "stem-permissive" conditions.
To attempt to identify the full set of genes involved in maintaining the stem cell state ("stemness"), high-throughput technologies like microarrays [ 13], chromatin immuno-precipitation [ 6] and MPSS [ 14] have been used to compare gene expression profiles between ES cells and differentiating cells.
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