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Continued monitoring of each of these wells over the second 4? days showed that 97% (279 out of 288) of the wells contained cells that continued to divide. Moreover, the range of times taken to complete the next division was the same as for the first division initiated in SF plus IL-11 plus UG26 CM, discounting the initial lag, and further addition of FBS plus SF plus IL-3 plus IL-6 plus Epo medium at the end of the second period of monitoring showed that 97% of the cultures again produced a very large clone over the following 7?days (Figure?6C). These results indicate that SFM containing SF plus IL-11 only is unable to support the survival of a large proportion of quiescent adult HSCs, although once activated, both their viability and their proliferation can be efficiently sustained http://www.selleckchem.com/Caspase.html by continued exposure to SF plus IL-11. However, this initial loss of quiescent adult BM ESLAM cells can be circumvented by exposure to UG26 CM or FBS plus IL-3 plus IL-6 plus Epo, although these two sources of prosurvival factors clearly differ in their mitogenic activities and in their abilities to sustain HSC self-renewal activity. Interestingly, the overall rate at which initially quiescent ESLAM BM cells enter the cell cycle appeared independent of any of these conditions of stimulation. As a first step toward Ceftiofur identifying the prosurvival factors produced by UG26 cells, we looked for pathways that they differentially activate as well as related candidate effectors secreted by UG26 cells and their possible receptors in purified adult BM ESLAM cells. To this end, we generated gene expression profiles for adult BM ESLAM cells before and 6?hr after being placed in culture with or without UG26 CM with or without SF plus IL-11. The 6?hr time point was chosen to obtain cells before evidence of their death is obvious when they are cultured in SF plus IL-11 alone (Lecault et?al., 2011). We then compared these profiles with each other, as well as with a published gene expression profile for UG26 cells (Ledran et?al., 2008) (Figure?7A). Analysis of the profiles obtained for ESLAM cells stimulated with UG26 CM (with or without SF plus IL-11) compared Ceritinib ic50 to fresh ESLAM cells and ESLAM cells stimulated with SF plus IL-11 alone identified a total of 250 (of 430 tested) REACTOME pathways for which some members showed significantly altered transcript expression (p?
