Wnt pathway modulation generates blastomere-derived mouse embryonic stem cells with different pluripotency features

Vila-Cejudo, Marta; Alonso-Alonso, Sandra; Pujol, Anna; Santaló, Josep; Ibáñez, Elena

doi:10.1007/s10815-020-01964-7

Cited by 4 publications

(3 citation statements)

References 42 publications

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“…Nonetheless, the derivation efficiency in R2i was slightly lower in our study (38.9% in the most efficient conditions) than the 46-50% obtained by Hassani et al (2014a). We tried to replicate the culture conditions from this other study by using N2B27 medium but, in agreement with previous results in our laboratory (Vila-Cejudo et al, 2019, 2020, we did not succeed in establishing Bm-ESC lines in this medium or did so but with extremely low efficiencies (<4%) both when using 2i as when using R2i. Thus, in our hands, ESC lines can only be efficiently established from single blastomeres of 8-cell embryos in KSR+LIF+2i or KSR+LIF+R2i.…”

Section: Discussionsupporting

confidence: 88%

Efficient generation of embryonic stem cells from single blastomeres of cryopreserved mouse embryos in the presence of signalling modulators

Alonso-Alonso

Santaló²,

Ibáñez³

2022

Reprod. Fertil. Dev.

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Context Derivation of embryonic stem cells (ESC) from single blastomeres is an interesting alternative to the use of whole blastocysts, but derivation rates are lower and the requirements for successful ESC obtention are still poorly defined. Aims To investigate the effects of embryo cryopreservation and of signalling modulators present during embryo culture and/or ESC establishment on ESC derivation efficiency from single 8-cell mouse blastomeres. Method Fresh and cryopreserved 2-cell embryos were cultured and biopsied at the 8-cell stage. Single blastomeres were cultured in the presence of 2i or R2i cocktails, with or without adrenocorticotropic hormone (ACTH). We analysed ESC derivation efficiencies and characterised pluripotency genes expression and karyotype integrity of the resulting lines. We also evaluated the impact of embryo preculture with R2i on epiblast cell numbers and derivation rates. Key results The ESC generation was not compromised by embryo cryopreservation and ACTH was dispensable under most of the conditions tested. While 2i and R2i were similarly effective for ESC derivation, R2i provided higher karyotype integrity. Embryo preculture with R2i yielded increased numbers of epiblast cells but did not lead to increased ESC generation. Conclusions Our findings help to define a simplified and efficient procedure for the establishment of mouse ESC from single 8-cell blastomeres. Implications This study will contribute to improving the potential of this experimental procedure, providing a tool to investigate the developmental potential of blastomeres isolated from different embryonic stages and to reduce the number of embryos needed for ESC derivation.

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Section: Discussionsupporting

confidence: 88%

Efficient generation of embryonic stem cells from single blastomeres of cryopreserved mouse embryos in the presence of signalling modulators

Alonso-Alonso

Santaló²,

Ibáñez³

2022

Reprod. Fertil. Dev.

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“…By analyzing the components of the induction medium, we fortuitously uncovered that Wnt/β-catenin signaling has a regulatory role. This relationship forced us to reconsider the function of CHIR99021 in iPSC and naïve ESC induction, wherein it is commonly used [ 28 , 29 ]. It is putative that Wnt/β-catenin signaling may be activated in the presence of CHIR99021 and therefore a low expression level of PRDM14.…”

Section: Discussionmentioning

confidence: 99%

PRDM14 extinction enables the initiation of trophoblast stem cell formation

Xu,

Zhao,

Peng

et al. 2024

Cell. Mol. Life Sci.

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Trophoblast stem cells (TSCs) can be chemically converted from embryonic stem cells (ESCs) in vitro. Although several transcription factors (TFs) have been recognized as essential for TSC formation, it remains unclear how differentiation cues link elimination of stemness with the establishment of TSC identity. Here, we show that PRDM14, a critical pluripotent circuitry component, is reduced during the formation of TSCs. The reduction is further shown to be due to the activation of Wnt/β-catenin signaling. The extinction of PRDM14 results in the erasure of H3K27me3 marks and chromatin opening in the gene loci of TSC TFs, including GATA3 and TFAP2C, which enables their expression and thus the initiation of the TSC formation process. Accordingly, PRDM14 reduction is proposed here as a critical event that couples elimination of stemness with the initiation of TSC formation. The present study provides novel insights into how induction signals initiate TSC formation.

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“…This could explain the extremely low rates of mESCs derivation from isolated blastomeres of 8-cell embryos ( Wakayama et al, 2007 ; González et al, 2010 ) and their partial rescue when incubating the blastomeres for 24 h with E-cadherin to mimic cell-cell contacts ( González et al, 2011b ). Similarly, the addition of 2i or R2i to the culture medium significantly improves mESCs derivation rates from single 8-cell blastomeres ( Hassani et al, 2014b ; Vila-Cejudo et al, 2020 ; Alonso-Alonso et al, 2022 ). The signaling modulators present in these cocktails have been shown to expand the pluripotent EPI population in intact embryos and isolated blastomeres ( Nichols et al, 2009 ; Van Der Jeught et al, 2014 ; Alonso-Alonso et al, 2022 ) and their effects on the maintenance of ESCs pluripotency are well known ( Ying et al, 2008 ).…”

Section: Discussionmentioning

confidence: 99%

Blastomeres of 8-cell mouse embryos differ in their ability to generate embryonic stem cells and produce lines with different transcriptional signatures

Alonso-Alonso,

Esteve-Codina,

Martin-Mur

et al. 2023

Front. Cell Dev. Biol.

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Embryonic stem cell (ESC) derivation from single blastomeres of 8-cell mouse embryos results in lower derivation rates than that from whole blastocysts, raising a biological question about the developmental potential of sister blastomeres. We aimed to assess the ability of 8-cell blastomeres to produce epiblast cells and ESC lines after isolation, and the properties of the resulting lines. Our results revealed unequal competence among sister blastomeres to produce ESC lines. At least half of the blastomeres possess a lower potential to generate ESCs, although culture conditions and blastomeres plasticity can redirect their non-pluripotent fate towards the epiblast lineage, allowing us to generate up to seven lines from the same embryo. Lines originated from the same embryo segregated into two groups according to their transcriptional signatures. While the expression of genes related to pluripotency and development was higher in one group, no differences were found in their trilineage differentiation ability. These results may help to improve our understanding of the ESC derivation process from single blastomeres and cell fate determination in the preimplantation mouse embryos.

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Wnt pathway modulation generates blastomere-derived mouse embryonic stem cells with different pluripotency features

Cited by 4 publications

References 42 publications

Efficient generation of embryonic stem cells from single blastomeres of cryopreserved mouse embryos in the presence of signalling modulators

Efficient generation of embryonic stem cells from single blastomeres of cryopreserved mouse embryos in the presence of signalling modulators

PRDM14 extinction enables the initiation of trophoblast stem cell formation

Blastomeres of 8-cell mouse embryos differ in their ability to generate embryonic stem cells and produce lines with different transcriptional signatures

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