Abstract
ABSTRACTMouse and rat embryonic stem cell (ESC) self-renewal can be maintained by dual inhibition of glycogen synthase kinase 3 (GSK3) and mitogen-activated protein kinase kinase (MEK). Inhibition of GSK3 promotes ESC self-renewal by abrogating T-cell factor 3 (TCF3)-mediated repression of the pluripotency network. How inhibition of MEK mediates ESC self-renewal, however, remains largely unknown. Here, we show that inhibition of MEK can significantly suppress lymphoid enhancer factor 1 (LEF1) expression in mouse ESCs. Knockdown or knockout of Lef1 partially mimics the self-renewal-promoting effect of MEK inhibitors. Moreover, depletion of both Tcf3 and Lef1 enables maintenance of undifferentiated mouse ESCs without exogenous factors, cytokines or inhibitors. Transcriptome resequencing analysis reveals that LEF1 is closely associated with endoderm specification in ESCs. Thus, our study adds support to the notion that the key to maintaining the ESC ground state is to shield ESCs from differentiative cues.
Highlights
Mouse embryonic stem cells are derived from preimplantation blastocysts and can be propagated extensively in culture while retaining the capacity to differentiate into all different cell types of the body (Evans and Kaufman, 1981; Huang et al, 2015; Martin, 1981)
We found that the self-renewal-promoting effect of PD03 in mESCs is partially attributable to the suppression of Lef1 expression and that depletion of Tcf3 and Lef1 can partially mimic the effect of 2i in maintaining embryonic stem cell (ESC) self-renewal
CHIR down-regulates T-cell factor 3 (TCF3) in mESCs Tcf3−/− mESC self-renewal could be maintained by PD03 alone (Fig. 1A,B), an outcome consistent with previous observations (Wray et al, 2011)
Summary
Mouse embryonic stem cells (mESCs) are derived from preimplantation blastocysts and can be propagated extensively in culture while retaining the capacity to differentiate into all different cell types of the body (Evans and Kaufman, 1981; Huang et al, 2015; Martin, 1981). We found that the self-renewal-promoting effect of PD03 in mESCs is partially attributable to the suppression of Lef1 expression and that depletion of Tcf3 and Lef1 can partially mimic the effect of 2i in maintaining ESC self-renewal. RESULTS AND DISSUSION CHIR down-regulates TCF3 in mESCs Tcf3−/− mESC self-renewal could be maintained by PD03 alone (Fig. 1A,B), an outcome consistent with previous observations (Wray et al, 2011).
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