scholarly journals Rapid and robust directed differentiation of mouse epiblast stem cells into definitive endoderm and forebrain organoids

2021 ◽  
Author(s):  
Daniel Medina-Cano ◽  
Emily K. Corrigan ◽  
Rachel A. Glenn ◽  
Mohammed Tarek Islam ◽  
Yuan Lin ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Development ◽  
Hippocampal Neurons ◽  
Embryonic Stem ◽  
Model System ◽  
Definitive Endoderm ◽  
Specific Cell ◽  
Directed Differentiation ◽  
Epiblast Stem Cells ◽  
Starting Point

Directed differentiation of pluripotent stem cells (PSCs) is a powerful model system for deconstructing embryonic development. Although mice are the most advanced mammalian model system for genetic studies of embryonic development, state-of-the-art protocols for directed differentiation of mouse PSCs into defined lineages tend to be slower and generate target cell types with lower purity than analogous protocols for human PSCs, limiting their application as models for mechanistic studies of development. Here, we examine the potential of mouse epiblast stem cells (EpiSCs) cultured in media containing Wnt pathway inhibitors (primed ground state conditions) as a starting point for directed differentiation. As a proof-of-concept, we focused our efforts on two specific cell/tissue types that have proven difficult to generate efficiently and reproducibly from mouse embryonic stem cells: definitive endoderm and neural organoids. First, we developed a new protocol that can rapidly generate nearly pure definitive endoderm from EpiSCs. Second, we developed a protocol for generating forebrain organoids that model the development of prethalamic and hippocampal neurons. These significantly improved differentiation models present new possibilities for combining mouse genetic tools and resources with in vitro differentiation to characterize the molecular and cellular mechanisms of embryonic development.

scholarly journals Early neurulation recapitulated in assemblies of embryonic and extraembryonic cells

2020 ◽  
Author(s):  
Noémie M. L. P. Bérenger-Currias ◽  
Maria Mircea ◽  
Esmée Adegeest ◽  
Patrick R. van den Berg ◽  
Marleen Feliksik ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Development ◽  
Neural Tube ◽  
Embryonic Stem ◽  
Specific Cell ◽  
Digestive Tube ◽  
Extraembryonic Endoderm ◽  
Starting Point

Recapitulating mammalian embryonic development in vitro is a major challenge in biology. It has been shown that gastruloids1–5 and ETX embryos6 can display hallmarks of gastrulation in vitro. However, these models fail to progress beyond spatially segregated, yet amorphous cellular assemblies. Systems such as organoids7 do show tissue stratification and organogenesis, but require adult stem cells or exogeneous induction of specific cell fates, and hence do not reflect the emergent organization of embryonic development. Notably, gastruloids are derived exclusively from embryonic stem cells (ESCs), whereas, in vivo, crucial patterning cues are provided by extraembryonic cells8. Here, we show that assemblies of mouse ESCs (mESCs) and extraembryonic endoderm (XEN) cells can develop beyond gastrulation and produce a central hallmark of organogenesis: stratified neural epithelia resembling a neural tube, which can be further differentiated to cerebral cortex-like tissue. By single-cell RNA-seq, we show that our model has a larger cell type diversity than existing models, and that mESCs and XEN cells impact each other’s differentiation. XEN cells promote neural tube formation through local inhibition of primitive streak formation. In turn, the presence of mESCs drives XEN cells to resemble visceral endoderm, which envelops the embryo in vivo. This study provides a model system to investigate neurulation and extraembryonic endoderm development, and may serve as a starting point to generate embryo models that advance further toward the formation of the vasculature, nervous system, and digestive tube.

scholarly journals Formative pluripotent stem cells show features of epiblast cells poised for gastrulation

Cell Research ◽  
2021 ◽  
Author(s):  
Xiaoxiao Wang ◽  
Yunlong Xiang ◽  
Yang Yu ◽  
Ran Wang ◽  
Yu Zhang ◽  
...  
Keyword(s):  
Stem Cells ◽  
Germ Cells ◽  
Pluripotent Stem Cells ◽  
Primordial Germ Cells ◽  
Embryonic Stem ◽  
Large Set ◽  
Mouse Escs ◽  
Epiblast Stem Cells ◽  
Early Gastrula

AbstractThe pluripotency of mammalian early and late epiblast could be recapitulated by naïve embryonic stem cells (ESCs) and primed epiblast stem cells (EpiSCs), respectively. However, these two states of pluripotency may not be sufficient to reflect the full complexity and developmental potency of the epiblast during mammalian early development. Here we report the establishment of self-renewing formative pluripotent stem cells (fPSCs) which manifest features of epiblast cells poised for gastrulation. fPSCs can be established from different mouse ESCs, pre-/early-gastrula epiblasts and induced PSCs. Similar to pre-/early-gastrula epiblasts, fPSCs show the transcriptomic features of formative pluripotency, which are distinct from naïve ESCs and primed EpiSCs. fPSCs show the unique epigenetic states of E6.5 epiblast, including the super-bivalency of a large set of developmental genes. Just like epiblast cells immediately before gastrulation, fPSCs can efficiently differentiate into three germ layers and primordial germ cells (PGCs) in vitro. Thus, fPSCs highlight the feasibility of using PSCs to explore the development of mammalian epiblast.

Germ line transmission of an inactive N-myc allele generated by homologous recombination in mouse embryonic stem cells

1990 ◽  
Vol 10 (12) ◽  
pp. 6755-6758
Author(s):  
B R Stanton ◽  
S W Reid ◽  
L F Parada
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Homologous Recombination ◽  
Embryonic Development ◽  
Cell Lines ◽  
Germ Line ◽  
Es Cells ◽  
Embryonic Stem ◽  
Es Cell ◽  
Germ Line Transmission

We have disrupted one allele of the N-myc locus in mouse embryonic stem (ES) cells by using homologous recombination techniques and have obtained germ line transmission of null N-myc ES cell lines with transmission of the null N-myc allele to the offspring. The creation of mice with a deficient N-myc allele will allow the generation of offspring bearing null N-myc alleles in both chromosomes and permit study of the role that this proto-oncogene plays in embryonic development.

scholarly journals Directed differentiation of telencephalic precursors from embryonic stem cells

2005 ◽  
Vol 8 (3) ◽  
pp. 288-296 ◽  
Author(s):  
Kiichi Watanabe ◽  
Daisuke Kamiya ◽  
Ayaka Nishiyama ◽  
Tomoko Katayama ◽  
Satoshi Nozaki ◽  
...  
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Directed Differentiation

scholarly journals Cellular Functions of OCT-3/4 Regulated by Ubiquitination in Proliferating Cells

Cancers ◽  
2020 ◽  
Vol 12 (3) ◽  
pp. 663
Author(s):  
Kwang-Hyun Baek ◽  
Jihye Choi ◽  
Chang-Zhu Pei
Keyword(s):  
Stem Cells ◽  
Cellular Proliferation ◽  
Critical Role ◽  
Embryonic Stem ◽  
Cell Types ◽  
Ubiquitin Proteasome System ◽  
Specific Cell ◽  
Cellular Mechanisms ◽  
Proliferating Cells ◽  
Proliferation And Differentiation

Octamer-binding transcription factor 3/4 (OCT-3/4), which is involved in the tumorigenesis of somatic cancers, has diverse functions during cancer development. Overexpression of OCT-3/4 has been detected in various human somatic tumors, indicating that OCT-3/4 activation may contribute to the development and progression of cancers. Stem cells can undergo self-renewal, pluripotency, and reprogramming with the help of at least four transcription factors, OCT-3/4, SRY box-containing gene 2 (SOX2), Krüppel-like factor 4 (KLF4), and c-MYC. Of these, OCT-3/4 plays a critical role in maintenance of undifferentiated state of embryonic stem cells (ESCs) and in production of induced pluripotent stem cells (iPSCs). Stem cells can undergo partitioning through mitosis and separate into specific cell types, three embryonic germ layers: the endoderm, the mesoderm, and the trophectoderm. It has been demonstrated that the stability of OCT-3/4 is mediated by the ubiquitin-proteasome system (UPS), which is one of the key cellular mechanisms for cellular homeostasis. The framework of the mechanism is simple, but the proteolytic machinery is complicated. Ubiquitination promotes protein degradation, and ubiquitination of OCT-3/4 leads to regulation of cellular proliferation and differentiation. Therefore, it is expected that OCT-3/4 may play a key role in proliferation and differentiation of proliferating cells.

Sign in / Sign up

Export Citation Format

Share Document