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Updated: May 8, 2026
Retroviral Infection of Murine Embryonic Stem Cell Derived Embryoid Body Cells for Analysis of Hematopoietic Differentiation
Published on: October 20, 2014
Identification of the missing pluripotency mediator downstream of leukaemia inhibitory factor
Graziano Martello1, Paul Bertone, Austin Smith
1Wellcome Trust-Medical Research Council Cambridge Stem Cell Institute, University of Cambridge, Cambridge, UK.
Abstract:
Self-renewal of pluripotent mouse embryonic stem (ES) cells is sustained by the cytokine leukaemia inhibitory factor (LIF) acting through the transcription factor Stat3. Several targets of Stat3 have previously been identified, most notably the reprogramming factor Klf4. However, such factors are neither required nor sufficient for the potent effect of LIF. We took advantage of Stat3 null ES cells to confirm that Stat3 mediates the self-renewal response to LIF. Through comparative transcriptome analysis intersected with genome location data, we arrived at a set of candidate transcription factor effectors. Among these, Tfcp2l1 (also known as Crtr-1) was most abundant. Constitutive expression of Tfcp2l1 at levels similar to those induced by LIF effectively substituted for LIF or Stat3 in sustaining clonal self-renewal and pluripotency. Conversely, knockdown of Tfcp2l1 profoundly compromised responsiveness to LIF. We further found that Tfcp2l1 is both necessary and sufficient to direct molecular reprogramming of post-implantation epiblast stem cells to naïve pluripotency. These results establish Tfcp2l1 as the principal bridge between LIF/Stat3 input and the transcription factor core of naïve pluripotency.
Insights
Leukaemia inhibitory factor (LIF) sustains mouse embryonic stem cell self-renewal via Stat3. Tfcp2l1 acts as the key transcription factor, bridging LIF signaling to pluripotency maintenance.
Area of Science:
- Stem cell biology
- Molecular genetics
- Epigenetics
Background:
- Pluripotent mouse embryonic stem (ES) cells require leukaemia inhibitory factor (LIF) for self-renewal, mediated by the transcription factor Stat3.
- While Stat3 targets like Klf4 are known, they don't fully explain LIF's potent effects.
Purpose of the Study:
- To identify the key transcription factors mediating LIF's role in ES cell self-renewal.
- To elucidate the molecular mechanism linking LIF/Stat3 signaling to pluripotency.
Main Methods:
- Comparative transcriptome analysis of Stat3 null ES cells.
- Genome location data integration to identify transcription factor targets.
- Tfcp2l1 expression manipulation (constitutive expression and knockdown).
Main Results:
- Tfcp2l1 was identified as the most abundant candidate transcription factor effector.
- Constitutive Tfcp2l1 expression mimicked LIF/Stat3 in sustaining self-renewal and pluripotency.
- Tfcp2l1 knockdown abolished LIF responsiveness.
- Tfcp2l1 was necessary and sufficient for reprogramming epiblast stem cells to naïve pluripotency.
Conclusions:
- Tfcp2l1 is the principal transcription factor linking LIF/Stat3 signaling to the core machinery of naïve pluripotency.
- Tfcp2l1 is a critical regulator of stem cell self-renewal and reprogramming.
