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The International Journal of... 2008The pancreas develops from two separate and independent endodermal primordia. The molecular events supporting the early morphological changes that give rise to the... (Review)
Review
The pancreas develops from two separate and independent endodermal primordia. The molecular events supporting the early morphological changes that give rise to the formation of the dorsal and ventral pancreatic buds result from coordinated responses to extrinsic and intrinsic signals. The extrinsic signals are involved in processes dictating whether progenitor cells remain as immature or as committed precursors. After specification, the sequential activation of transcription factors determines cell autonomously the commitment and differentiation of these progenitors. During pancreas development, the roles of extrinsic and intrinsic signals are variable, depending on the particular competence of each progenitor cell. We summarize in this review the main events, at the level of gene expression, which are involved in the early stages of pancreas development.
Topics: Animals; Cell Differentiation; Cell Lineage; Embryo, Mammalian; Endoderm; Gene Expression Regulation, Developmental; Homeodomain Proteins; Islets of Langerhans; Mesoderm; Organogenesis; Pancreas; Signal Transduction; Stem Cells; Trans-Activators; Transcription Factors
PubMed: 18956314
DOI: 10.1387/ijdb.072444cb -
Journal of Molecular Cell Biology Aug 2020Recent advances in development of protocols for directed differentiation from human pluripotent stem cells (hPSCs) to defined lineages, in combination with 3D organoid... (Review)
Review
Recent advances in development of protocols for directed differentiation from human pluripotent stem cells (hPSCs) to defined lineages, in combination with 3D organoid technology, have facilitated the generation of various endoderm-derived organoids for in vitro modeling of human gastrointestinal development and associated diseases. In this review, we discuss current state-of-the-art strategies for generating hPSC-derived endodermal organoids including stomach, liver, pancreatic, small intestine, and colonic organoids. We also review the advantages of using this system to model various human diseases and evaluate the shortcomings of this technology. Finally, we emphasize how other technologies, such as genome editing and bioengineering, can be incorporated into the 3D hPSC-organoid models to generate even more robust and powerful platforms for understanding human organ development and disease modeling.
Topics: Disease; Endoderm; Humans; Models, Biological; Organogenesis; Organoids; Pluripotent Stem Cells
PubMed: 32652003
DOI: 10.1093/jmcb/mjaa031 -
Current Opinion in Genetics &... Aug 2013Ventral folding morphogenesis (VFM), a vital morphogenetic process in amniotes, mediates gut endoderm internalization, linear heart tube formation, ventral body wall... (Review)
Review
Ventral folding morphogenesis (VFM), a vital morphogenetic process in amniotes, mediates gut endoderm internalization, linear heart tube formation, ventral body wall closure and encasement of the fetus in extraembryonic membranes. Aberrant VFM underlies a number of birth defects such as gastroschisis and ectopia cordis in human and misplacement of head and heart in mouse. Recent cell lineage-specific mouse mutant analyses identified the Bone Morphogenetic Protein (BMP) pathway and Anterior Visceral Endoderm (AVE) as key regulators of anterior VFM. Loss of BMP2 expression solely from embryonic visceral endoderm (EmVE) and the AVE blocks formation of foregut invagination, and simultaneously, aberrantly positions the heart anterior/dorsal to the head, suggesting a mechanistic link between foregut and head/heart morphogenesis.
Topics: Animals; Body Patterning; Bone Morphogenetic Protein 2; Cell Lineage; Embryonic Development; Endoderm; Gene Expression Regulation, Developmental; Head; Heart; Humans; Mice; Morphogenesis
PubMed: 23706163
DOI: 10.1016/j.gde.2013.04.001 -
Cell Stem Cell Apr 2012The lung is composed of numerous epithelial lineages that arise from the anterior foregut endoderm. This review discusses how insights into the signaling mechanisms that... (Review)
Review
The lung is composed of numerous epithelial lineages that arise from the anterior foregut endoderm. This review discusses how insights into the signaling mechanisms that regulate lung endoderm specification and subsequent differentiation have recently been exploited to direct differentiation of hESCs/iPSCs into expandable lung progenitors.
Topics: Animals; Cell Differentiation; Endoderm; Humans; Lung; Pluripotent Stem Cells; Signal Transduction
PubMed: 22482501
DOI: 10.1016/j.stem.2012.03.013 -
Developmental Biology Apr 2007In spite of the therapeutic importance of endoderm derivatives such as the pancreas, liver, lung, and intestine, there are few molecular markers specific for early...
In spite of the therapeutic importance of endoderm derivatives such as the pancreas, liver, lung, and intestine, there are few molecular markers specific for early endoderm. In order to identify endoderm-specific genes as well as to define transcriptional differences between definitive and visceral endoderm, we performed microarray analysis on E8.25 definitive and visceral endoderm. We have developed an early endoderm gene expression signature, and clarified the transcriptional similarities and differences between definitive and visceral endoderm. Additionally, we have developed methods for flow cytometric isolation of definitive and visceral endoderm. These results shed light on the mechanism of endoderm formation and should facilitate investigation of endoderm formation from embryonic stem cells.
Topics: Animals; Cell Differentiation; Cells, Cultured; Embryonic Stem Cells; Endoderm; Gene Expression; Mesoderm; Mice; Mice, Transgenic; Multipotent Stem Cells; Oligonucleotide Array Sequence Analysis; Viscera
PubMed: 17328885
DOI: 10.1016/j.ydbio.2007.01.011 -
Genes & Development Aug 2008The major forms of diabetes are characterized by pancreatic islet beta-cell dysfunction and decreased beta-cell numbers, raising hope for cell replacement therapy.... (Review)
Review
The major forms of diabetes are characterized by pancreatic islet beta-cell dysfunction and decreased beta-cell numbers, raising hope for cell replacement therapy. Although human islet transplantation is a cell-based therapy under clinical investigation for the treatment of type 1 diabetes, the limited availability of human cadaveric islets for transplantation will preclude its widespread therapeutic application. The result has been an intense focus on the development of alternate sources of beta cells, such as through the guided differentiation of stem or precursor cell populations or the transdifferentiation of more plentiful mature cell populations. Realizing the potential for cell-based therapies, however, requires a thorough understanding of pancreas development and beta-cell formation. Pancreas development is coordinated by a complex interplay of signaling pathways and transcription factors that determine early pancreatic specification as well as the later differentiation of exocrine and endocrine lineages. This review describes the current knowledge of these factors as they relate specifically to the emergence of endocrine beta cells from pancreatic endoderm. Current therapeutic efforts to generate insulin-producing beta-like cells from embryonic stem cells have already capitalized on recent advances in our understanding of the embryonic signals and transcription factors that dictate lineage specification and will most certainly be further enhanced by a continuing emphasis on the identification of novel factors and regulatory relationships.
Topics: Animals; Cell Differentiation; Cell Lineage; Embryo, Mammalian; Embryonic Stem Cells; Endoderm; Humans; Insulin-Secreting Cells; Models, Biological; Organogenesis; Pancreas; Signal Transduction; Transcription, Genetic
PubMed: 18676806
DOI: 10.1101/gad.1670808 -
Cell Reports Dec 2020The ever-increasing therapeutic and pharmaceutical demand for liver cells calls for systems that enable mass production of hepatic cells. Here we describe a large-scale...
The ever-increasing therapeutic and pharmaceutical demand for liver cells calls for systems that enable mass production of hepatic cells. Here we describe a large-scale suspension system that uses human endoderm stem cells (hEnSCs) as precursors to generate functional and transplantable hepatocytes (E-heps) or cholangiocytes (E-chos). hEnSC-derived hepatic populations are characterized by single-cell transcriptomic analyses and compared with hESC-derived counterparts, in-vitro-maintained or -expanded primary hepatocytes and adult cells, which reveals that hepatic differentiation of hEnSCs recapitulates in vivo development and that the heterogeneities of the resultant populations can be manipulated by regulating the EGF and MAPK signaling pathways. Functional assessments demonstrate that E-heps and E-chos possess properties comparable with adult counterparts and that, when transplanted intraperitoneally, encapsulated E-heps were able to rescue rats with acute liver failure. Our study lays the foundation for cell-based therapeutic agents and in vitro applications for liver diseases.
Topics: Bile Ducts; Cell Culture Techniques; Cell Differentiation; Endoderm; Epithelial Cells; Hepatocytes; Human Embryonic Stem Cells; Humans; Liver; Pluripotent Stem Cells
PubMed: 33296648
DOI: 10.1016/j.celrep.2020.108455 -
Nature Communications Jan 2023The correct establishment of DNA methylation patterns during mouse early development is essential for cell fate specification. However, the molecular targets as well as...
The correct establishment of DNA methylation patterns during mouse early development is essential for cell fate specification. However, the molecular targets as well as the mechanisms that determine the specificity of the de novo methylation machinery during differentiation are not completely elucidated. Here we show that the DNMT3B-dependent DNA methylation of key developmental regulatory regions at epiblast-like cells (EpiLCs) provides an epigenetic priming that ensures flawless commitment at later stages. Using in vitro stem cell differentiation and loss of function experiments combined with high-throughput genome-wide bisulfite-, bulk-, and single cell RNA-sequencing we dissected the specific role of DNMT3B in cell fate. We identify DNMT3B-dependent regulatory elements on the genome which, in Dnmt3b knockout (3BKO), impair the differentiation into meso-endodermal (ME) progenitors and redirect EpiLCs towards the neuro-ectodermal lineages. Moreover, ectopic expression of DNMT3B in 3BKO re-establishes the DNA methylation of the master regulator Sox2 super-enhancer, downmodulates its expression, and restores the expression of ME markers. Taken together, our data reveal that DNMT3B-dependent methylation at the epiblast stage is essential for the priming of the meso-endodermal lineages and provide functional characterization of the de novo DNMTs during EpiLCs lineage determination.
Topics: Animals; Mice; Mouse Embryonic Stem Cells; Endoderm; DNA (Cytosine-5-)-Methyltransferases; Cell Differentiation; Cell Lineage; DNA Methylation
PubMed: 36690616
DOI: 10.1038/s41467-023-35938-x -
Philosophical Transactions of the Royal... Dec 2014Mouse epiblast stem cells (EpiSCs) display temporal differences in the upregulation of Mixl1 expression during the initial steps of in vitro differentiation, which can... (Review)
Review
Mouse epiblast stem cells (EpiSCs) display temporal differences in the upregulation of Mixl1 expression during the initial steps of in vitro differentiation, which can be correlated with their propensity for endoderm differentiation. EpiSCs that upregulated Mixl1 rapidly during differentiation responded robustly to both Activin A and Nodal in generating foregut endoderm and precursors of pancreatic and hepatic tissues. By contrast, EpiSCs that delayed Mixl1 upregulation responded less effectively to Nodal and showed an overall suboptimal outcome of directed differentiation. The enhancement in endoderm potency in Mixl1-early cells may be accounted for by a rapid exit from the progenitor state and the efficient response to the induction of differentiation by Nodal. EpiSCs that readily differentiate into the endoderm cells are marked by a distinctive expression fingerprint of transforming growth factor (TGF)-β signalling pathway genes and genes related to the endoderm lineage. Nodal appears to elicit responses that are associated with transition to a mesenchymal phenotype, whereas Activin A promotes gene expression associated with maintenance of an epithelial phenotype. We postulate that the formation of definitive endoderm (DE) in embryoid bodies follows a similar process to germ layer formation from the epiblast, requiring an initial de-epithelialization event and subsequent re-epithelialization. Our results show that priming EpiSCs with the appropriate form of TGF-β signalling at the formative phase of endoderm differentiation impacts on the further progression into mature DE-derived lineages, and that this is influenced by the initial characteristics of the cell population. Our study also highlights that Activin A, which is commonly used as an in vitro surrogate for Nodal in differentiation protocols, does not elicit the same downstream effects as Nodal, and therefore may not effectively mimic events that take place in the mouse embryo.
Topics: Animals; Cell Differentiation; Embryonic Stem Cells; Endoderm; Gene Expression Regulation, Developmental; Germ Layers; Homeodomain Proteins; Inhibin-beta Subunits; Mice; Nodal Protein; Signal Transduction; Transforming Growth Factor beta
PubMed: 25349457
DOI: 10.1098/rstb.2013.0550 -
Developmental Cell Jan 2018Specification of the three germ layers by graded Nodal signaling has long been seen as a paradigm for patterning through a single morphogen gradient. However, by...
Specification of the three germ layers by graded Nodal signaling has long been seen as a paradigm for patterning through a single morphogen gradient. However, by exploiting the unique properties of the zebrafish embryo to capture the dynamics of signaling and cell fate allocation, we now demonstrate that Nodal functions in an incoherent feedforward loop, together with Fgf, to determine the pattern of endoderm and mesoderm specification. We show that Nodal induces long-range Fgf signaling while simultaneously inducing the cell-autonomous Fgf signaling inhibitor Dusp4 within the first two cell tiers from the margin. The consequent attenuation of Fgf signaling in these cells allows specification of endoderm progenitors, while the cells further from the margin, which receive Nodal and/or Fgf signaling, are specified as mesoderm. This elegant model demonstrates the necessity of feedforward and feedback interactions between multiple signaling pathways for providing cells with temporal and positional information.
Topics: Animals; Dual-Specificity Phosphatases; Endoderm; Extracellular Signal-Regulated MAP Kinases; Feedback, Physiological; Fibroblast Growth Factors; MAP Kinase Signaling System; Mesoderm; Nodal Signaling Ligands; Zebrafish; Zebrafish Proteins
PubMed: 29275993
DOI: 10.1016/j.devcel.2017.11.021