Mouse Embryonic Stem Cells

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Cat.No.

CSC-00864L

Description

Mouse embryonic stem cells (mESCs) were first isolated and propagated in culture in 1981. mESCs are typically isolated from blastocysts from the inner cell mass of 3.5-day-old embryos and have the potential to generate every cell type found in the body. They can be maintained in an undifferentiated, pluripotent state by culture with leukemia inhibitory factor (LIF) on a feeder layer of mitotically arrested mouse embryonic fibroblasts or in a feeder-free environment using gelatin-coated flasks.

mESCs are invaluable in understanding many aspects of cell and developmental biology, including cell cycle regulation, cellular interactions during development, and the control of gene expression. mESCs will continue to be an important system for understanding stem cell and tissue biology and for creating transgenic or knock-out mice, used as in vivo disease models, by homologous recombination. Recent applications of mESCs include identifying factors that maintain pluripotency, human genetic disease research, identifying novel differentiation factors, and for cancer biology.

Species

Mouse

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For Research Use Only

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Dry Ice

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Mouse embryonic stem cells (mESCs) are pluripotent cell lines derived from the inner cell mass (ICM) of preimplantation blastocysts, typically at embryonic day 3.5 (E3.5) in the C57BL/6 or 129/Sv strain backgrounds. mESCs exhibit unlimited self-renewal capacity while retaining the potential to differentiate into all three germ layers-ectoderm, mesoderm, and endoderm-both in vitro and in vivo. Under permissive culture conditions supplemented with leukemia inhibitory factor (LIF) and bone morphogenetic protein 4 (BMP4), or in ground-state conditions using dual inhibition (2i) of MEK and GSK3β, mESCs maintain pluripotency with minimal spontaneous differentiation. They express core pluripotency markers including OCT4, SOX2, NANOG, and SSEA-1, while lacking the differentiation-associated marker SSEA-3.

The unparalleled advantages of mESCs lie in their superior genetic tractability and biological fidelity. These cells support high-efficiency homologous recombination, enabling targeted gene knockouts, knock-ins, and conditional alleles-pioneering the creation of thousands of transgenic mouse models. Their ability to contribute to germline chimeras upon blastocyst injection remains the definitive functional assay for pluripotency. Additionally, mESCs are amenable to CRISPR/Cas9-mediated genome editing, base editing, and epigenome engineering with remarkable ease. In vitro differentiation protocols can recapitulate early embryonic developmental processes, including gastrulation, neurogenesis, and cardiogenesis, making mESCs a powerful system for dissecting signaling pathways and epigenetic regulatory networks. Furthermore, mESCs serve as a robust platform for toxicological screening, drug discovery, and the development of cell-based therapies. Their rapid growth, clonal stability, and well-established culture protocols render them the gold-standard pluripotent stem cell model. Despite the emergence of human iPSCs, mESCs remain indispensable for fundamental developmental biology and preclinical translational research.

Genome-Wide Silencer Screening Reveals Key Silencer Modulating Reprogramming Efficiency in Mouse Induced Pluripotent Stem Cells

The majority of the mouse genome is composed of non-coding regions, which harbor numerous regulatory sequences essential for gene regulation. While extensive research focuses on enhancers that activate gene expression, the role of silencers that repress gene expression remains less explored. In this study, the first genome-wide identification of silencers in the mouse genome is conducted.

In mouse embryonic fibroblasts (MEFs) and embryonic stem cells (mESCs), 89 596 and 115 165 silencers are identified, respectively. These silencers are ubiquitously distributed across the genome and are predominantly associated with low-expression genes. Additionally, these silencers are mainly cell-specific and function by binding to repressive transcription factors (TFs). Further, these silencers are notably enriched with the histone modification H3K9me3. It is observed that the transformation between dual-function silencers and enhancers is correlated with intracellular transcription factor concentrations, accompanied by changes in epigenetic modifications. In terms of biological effects, we have identified silencers that can enhance the induction efficiency of MEFs and influence the pluripotency of mESCs.

Collectively, this work offers the first comprehensive silencer landscape in the mouse genome and provides strong evidence for the role of silencers in the induction of induced pluripotent stem cells (iPSCs).

Knockout of Smad2 Silencers Promotes Differentiation of Mouse Embryonic Stem Cells. — Fig. 1. SS1 and SS2 silencers regulate *Smad2* gene expression and pluripotency in mESCs (Zhu, Xiusheng, *et al.*, 2025).

Knockout of Nanog Silencer Significantly Enhances iPSCs Induction Efficiency. — Fig. 2. Knockout of *Nanog* silencers significantly increases the induction efficiency of iPSCs (Zhu, Xiusheng, *et al.*, 2025).

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For research use only. Not for any other purpose.