The murine H19 gene is activated during embryonic stem cell differentiation in vitro and at the time of implantation in the developing embryo

Development ◽  
1991 ◽  
Vol 113 (4) ◽  
pp. 1105-1114 ◽  
Author(s):  
F. Poirier ◽  
C.T. Chan ◽  
P.M. Timmons ◽  
E.J. Robertson ◽  
M.J. Evans ◽  
...  
Keyword(s):  
Embryonic Stem ◽  
Stem Cell Differentiation ◽  
Cell Types ◽  
Embryoid Bodies ◽  
Embryonic Stem Cell Differentiation ◽  
Cdna Clones ◽  
Stage Of Development ◽  
Before And After

The differentiation in vitro of murine embryonic stem cells to embryoid bodies mimics events that occur in vivo shortly before and after embryonic implantation. We have used this system, together with differential cDNA cloning, to identify genes the expression of which is regulated during early embryogenesis. Here we describe the isolation of several such cDNA clones, one of which corresponds to the gene H19. This gene is activated in extraembryonic cell types at the time of implantation, suggesting that it may play a role at this stage of development, and is subsequently expressed in all of the cells of the mid-gestation embryo with the striking exception of most of those of the developing central and peripheral nervous systems. After birth, expression of this gene ceases or is dramatically reduced in all tissues.

scholarly journals Extracellular Matrix and Integrins in Embryonic Stem Cell Differentiation

2015 ◽  
Vol 8s2 ◽  
pp. BCI.S30377 ◽  
Author(s):  
Han Wang ◽  
Xie Luo ◽  
Jake Leighton
Keyword(s):  
Extracellular Matrix ◽  
Pancreatic Beta Cells ◽  
Embryonic Stem ◽  
Stem Cell Differentiation ◽  
Embryonic Stem Cell Differentiation ◽  
In Vitro Differentiation ◽  
Integrin Receptors

Embryonic stem cells (ESCs) are pluripotent cells with great therapeutic potentials. The in vitro differentiation of ESC was designed by recapitulating embryogenesis. Significant progress has been made to improve the in vitro differentiation protocols by toning soluble maintenance factors. However, more robust methods for lineage-specific differentiation and maturation are still under development. Considering the complexity of in vivo embryogenesis environment, extracellular matrix (ECM) cues should be considered besides growth factor cues. ECM proteins bind to cells and act as ligands of integrin receptors on cell surfaces. Here, we summarize the role of the ECM and integrins in the formation of three germ layer progenies. Various ECM–integrin interactions were found, facilitating differentiation toward definitive endoderm, hepatocyte-like cells, pancreatic beta cells, early mesodermal progenitors, cardiomyocytes, neuroectoderm lineages, and epidermal cells, such as keratinocytes and melanocytes. In the future, ECM combinations for the optimal ESC differentiation environment will require substantial study.

Specific Effect of 5-Fluorouracil on α-Fetoprotein Gene Expression During the In Vitro Mouse Embryonic Stem Cell Differentiation

2010 ◽  
Vol 29 (3) ◽  
pp. 297-304 ◽  
Author(s):  
David Pamies ◽  
Néstor Vicente-Salar ◽  
Miguel A. Sogorb ◽  
Enrique Roche ◽  
Juan A. Reig
Keyword(s):  
Es Cells ◽  
Embryonic Stem ◽  
Stem Cell Differentiation ◽  
Embryoid Bodies ◽  
Screening Methods ◽  
Embryonic Stem Cell Differentiation ◽  
Visceral Endoderm ◽  
Gene Markers ◽  
Wide Range

Embryonic stem (ES) cells are considered an important alternative to develop in vitro screening methods for embryotoxicity. Mouse ES cells can be cultured as cell suspension aggregates termed “embryoid bodies” (EBs) in which cells start to differentiate. We have studied the expression of several genes in the presence of a wide range of concentrations of 5-fluorouracil (5-FU). This well-established embryotoxic compound completely inhibited cell viability at 200 nmol/L in monolayer cultures. At lower concentrations, 5-FU led to decrease in the expression of the α-fetoprotein gene, a marker of the visceral endoderm, in the EBs. However, the expression of several mesodermal gene markers was not significantly affected at these concentrations. These results suggest a high sensitivity of the visceral endoderm differentiation to 5-FU. Therefore, the quantification of the α-fetoprotein gene after exposure to potential embryotoxicants should be considered an additional end point in future embryotoxicity assays in vitro with ES cells.

scholarly journals Mouse lysocardiolipin acyltransferase controls the development of hematopoietic and endothelial lineages during in vitro embryonic stem-cell differentiation

Blood ◽  
2007 ◽  
Vol 110 (10) ◽  
pp. 3601-3609 ◽  
Author(s):  
Chengyan Wang ◽  
Patrick W. Faloon ◽  
Zhijia Tan ◽  
Yaxin Lv ◽  
Pengbo Zhang ◽  
...  
Keyword(s):  
Cell Differentiation ◽  
Messenger Rna ◽  
Molecular Mechanisms ◽  
Embryonic Stem ◽  
Stem Cell Differentiation ◽  
Embryoid Bodies ◽  
Embryonic Stem Cell Differentiation ◽  
Hematopoietic Stem ◽  
Endothelial Genes

Abstract The blast colony-forming cell (BL-CFC) was identified as an equivalent to the hemangioblast during in vitro embryonic stem (ES) cell differentiation. However, the molecular mechanisms underlying the generation of the BL-CFC remain largely unknown. Here we report the isolation of mouse lysocardiolipin acyltransferase (Lycat) based on homology to zebrafish lycat, a candidate gene for the cloche locus. Mouse Lycat is expressed in hematopoietic organs and is enriched in the Lin−C-Kit+Sca-1+ hematopoietic stem cells in bone marrow and in the Flk1+/hCD4+(Scl+) hemangioblast population in embryoid bodies. The forced Lycat transgene leads to increased messenger RNA expression of hematopoietic and endothelial genes as well as increased blast colonies and their progenies, endothelial and hematopoietic lineages. The Lycat small interfering RNA transgene leads to a decrease expression of hematopoietic and endothelial genes. An unbiased genomewide microarray analysis further substantiates that the forced Lycat transgene specifically up-regulates a set of genes related to hemangioblasts and hematopoietic and endothelial lineages. Therefore, mouse Lycat plays an important role in the early specification of hematopoietic and endothelial cells, probably acting at the level of the hemangioblast.

scholarly journals Defining the earliest step of cardiovascular progenitor specification during embryonic stem cell differentiation

2011 ◽  
Vol 192 (5) ◽  
pp. 751-765 ◽  
Author(s):  
Antoine Bondue ◽  
Simon Tännler ◽  
Giuseppe Chiapparo ◽  
Samira Chabab ◽  
Mirana Ramialison ◽  
...  
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Fluorescent Protein ◽  
Transcriptional Profiling ◽  
Embryonic Stem ◽  
Stem Cell Differentiation ◽  
Embryonic Stem Cell Differentiation ◽  
Heart Fields

During embryonic development and embryonic stem cell (ESC) differentiation, the different cell lineages of the mature heart arise from two types of multipotent cardiovascular progenitors (MCPs), the first and second heart fields. A key question is whether these two MCP populations arise from differentiation of a common progenitor. In this paper, we engineered Mesp1–green fluorescent protein (GFP) ESCs to isolate early MCPs during ESC differentiation. Mesp1-GFP cells are strongly enriched for MCPs, presenting the ability to differentiate into multiple cardiovascular lineages from both heart fields in vitro and in vivo. Transcriptional profiling of Mesp1-GFP cells uncovered cell surface markers expressed by MCPs allowing their prospective isolation. Mesp1 is required for MCP specification and the expression of key cardiovascular transcription factors. Isl1 is expressed in a subset of early Mesp1-expressing cells independently of Mesp1 and acts together with Mesp1 to promote cardiovascular differentiation. Our study identifies the early MCPs residing at the top of the cellular hierarchy of cardiovascular lineages during ESC differentiation.

Na+/H+ exchanger isoform 1 facilitates cardiomyocyte embryonic stem cell differentiation

2009 ◽  
Vol 296 (1) ◽  
pp. H159-H170 ◽  
Author(s):  
Xiuju Li ◽  
Pratap Karki ◽  
Lei Lei ◽  
Huayan Wang ◽  
Larry Fliegel
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Cell Differentiation ◽  
Embryonic Stem ◽  
Stem Cell Differentiation ◽  
Embryoid Bodies ◽  
Embryonic Stem Cell Differentiation ◽  
Cardiomyocyte Differentiation ◽  
Nhe1 Activity

Embryonic stem cells provide one potential source of cardiomyocytes for cardiac transplantation; however, after differentiation of stem cells in vitro, cardiomyocytes usually account for only a minority of cells present. To gain insights into improving cardiomyocyte development from stem cells, we examined the role of the Na+/H+ exchanger isoform 1 (NHE1) in cardiomyocyte differentiation. NHE1 protein and message levels were induced by treatment of CGR8 cells to form embryoid bodies and cardiomyocytes. The NHE1 protein was present on the cell surface and NHE1 inhibitor-sensitive activity was detected. Inhibition of NHE1 activity during differentiation of CGR8 cells prevented cardiomyocyte differentiation as indicated by decreased message for transcription factors Nkx2-5 and Tbx5 and decreased levels of α-myosin heavy chain protein. Increased expression of NHE1 from an adenoviral vector facilitated cardiomyocyte differentiation. Similar results were found with cardiomyocyte differentiation of P19 embryonal carcinoma cells. CGR8 cells were treated to induce differentiation, but when differentiation was inhibited by dispersing the EBs, myocardial development was inhibited. The results demonstrate that NHE1 activity is important in facilitating stem cell differentiation to cardiomyocyte lineage. Elevated NHE1 expression appears to be triggered as part of the process that facilitates cardiomyocyte development.

scholarly journals Convergence of Notch and β-catenin signaling induces arterial fate in vascular progenitors

2010 ◽  
Vol 189 (2) ◽  
pp. 325-338 ◽  
Author(s):  
Kohei Yamamizu ◽  
Taichi Matsunaga ◽  
Hideki Uosaki ◽  
Hiroyuki Fukushima ◽  
Shiori Katayama ◽  
...  
Keyword(s):  
Molecular Mechanisms ◽  
Embryonic Stem ◽  
Gene Promoter ◽  
Stem Cell Differentiation ◽  
Embryonic Stem Cell Differentiation ◽  
Target Gene Promoter ◽  
Vascular Progenitor Cells ◽  
Molecular Machinery

Molecular mechanisms controlling arterial–venous specification have not been fully elucidated. Previously, we established an embryonic stem cell differentiation system and demonstrated that activation of cAMP signaling together with VEGF induces arterial endothelial cells (ECs) from Flk1+ vascular progenitor cells. Here, we show novel arterial specification machinery regulated by Notch and β-catenin signaling. Notch and GSK3β-mediated β-catenin signaling were activated downstream of cAMP through phosphatidylinositol-3 kinase. Forced activation of Notch and β-catenin with VEGF completely reconstituted cAMP-elicited arterial EC induction, and synergistically enhanced target gene promoter activity in vitro and arterial gene expression during in vivo angiogenesis. A protein complex with RBP-J, the intracellular domain of Notch, and β-catenin was formed on RBP-J binding sites of arterial genes in arterial, but not venous ECs. This molecular machinery for arterial specification leads to an integrated and more comprehensive understanding of vascular signaling.

scholarly journals Effects of early geometric confinement on the transcriptomic profile of human cerebral organoids

2021 ◽  
Author(s):  
Dilara Sen ◽  
Alexis Voulgaropoulos ◽  
Albert J. Keung
Keyword(s):  
Stem Cell Differentiation ◽  
Cell Types ◽  
Brain Regions ◽  
Embryoid Bodies ◽  
Model Systems ◽  
Human Model ◽  
Spatial Factors ◽  
Neural Lineage

ABSTRACTBackgroundBiophysical factors such as shape and mechanical forces are known to play crucial roles in stem cell differentiation, embryogenesis and neurodevelopment. However, the complexity and experimental challenges capturing such early stages of development, and ethical concerns associated with human embryo and fetal research, limit our understanding of how these factors affect human brain organogenesis. Human cerebral organoids (hCO) are attractive models due to their ability to model important brain regions and transcriptomics of early in vivo brain development. Furthermore, they provide three-dimensional environments that better mimic the in vivo environment. To date, they have been used to understand the effects of genetics and soluble factors on neurodevelopment. Establishing links between spatial factors and hCO development will require the development of new approaches.ResultsHere, we investigated the effects of early geometric confinements on transcriptomic changes during hCO differentiation. Using a custom and tunable agarose microwell platform we generated embryoid bodies (EB) of diverse shapes and then further differentiated those EBs to whole brain hCOs. Our results showed that the microwells did not have negative gross impacts on the ability of the hCOs to differentiate generally towards neural fates, and there were clear shape dependent effects on neural lineage specification. In particular, we observed that non-spherical shapes showed signs of altered neurodevelopmental kinetics and favored the development of medial ganglionic eminence-associated brain regions and cell types over cortical regions.ConclusionsThe findings presented here suggest a role for spatial factors in brain region specification during hCO development. Understanding these spatial patterning factors will not only improve understanding of in vivo development and differentiation, but also provide important handles with which to advance and improve control over human model systems for in vitro applications.

scholarly journals Diverse but unique astrocytic phenotypes during embryonic stem cell differentiation, culturing and aging

2021 ◽  
Author(s):  
Kiara Freitag ◽  
Pascale Eede ◽  
Andranik Ivanov ◽  
Shirin Schneeberger ◽  
Tatiana Borodina ◽  
...  
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Embryonic Stem ◽  
Calcium Signalling ◽  
Stem Cell Differentiation ◽  
Embryonic Stem Cell Differentiation ◽  
The Central Nervous System ◽  
Transcriptomic Level

Astrocytes are resident glia cells of the central nervous system (CNS) that play complex and heterogeneous roles in brain development, homeostasis and disease. Since their vast involvement in health and disease is becoming increasingly recognized, suitable and reliable tools for studying these cells in vivo and in vitro are of utmost importance. One of the key challenges hereby is to adequately mimic their context-dependent in vivo phenotypes and functions in vitro. To better understand the spectrum of astrocytic variations in defined settings we performed a side-by-side-comparison of embryonic stem cell (ESC)-derived astrocytes as well as primary neonatal and adult astrocytes, revealing major differences on a functional and transcriptomic level, specifically on proliferation, migration, calcium signalling and cilium activity. Our results highlight the need to carefully consider the choice of astrocyte origin and phenotype with respect to age, isolation and culture protocols based on the respective biological question.

scholarly journals The Lysine Methylase SMYD3 Modulates Mesendodermal Commitment during Development

Cells ◽  
2021 ◽  
Vol 10 (5) ◽  
pp. 1233
Author(s):  
Raffaella Fittipaldi ◽  
Pamela Floris ◽  
Valentina Proserpio ◽  
Franco Cotelli ◽  
Monica Beltrame ◽  
...  
Keyword(s):  
Embryonic Stem ◽  
Stem Cell Differentiation ◽  
Model Systems ◽  
Embryonic Stem Cell Differentiation ◽  
In Vitro Differentiation ◽  
Cancer Models ◽  
Lineage Markers

SMYD3 (SET and MYND domain containing protein 3) is a methylase over-expressed in cancer cells and involved in oncogenesis. While several studies uncovered key functions for SMYD3 in cancer models, the SMYD3 role in physiological conditions has not been fully elucidated yet. Here, we dissect the role of SMYD3 at early stages of development, employing mouse embryonic stem cells (ESCs) and zebrafish as model systems. We report that SMYD3 depletion promotes the induction of the mesodermal pattern during in vitro differentiation of ESCs and is linked to an upregulation of cardiovascular lineage markers at later stages. In vivo, smyd3 knockdown in zebrafish favors the upregulation of mesendodermal markers during zebrafish gastrulation. Overall, our study reveals that SMYD3 modulates levels of mesendodermal markers, both in development and in embryonic stem cell differentiation.

scholarly journals The dynamic emergence of GATA1 complexes identified in in vitro embryonic stem cell differentiation and in vivo mouse fetal liver

Haematologica ◽  
2019 ◽  
Vol 105 (7) ◽  
pp. 1802-1812
Author(s):  
Xiao Yu ◽  
Andrea Martella ◽  
Petros Kolovos ◽  
Mary Stevens ◽  
Ralph Stadhouders ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Differentiation ◽  
Embryonic Stem Cell ◽  
Fetal Liver ◽  
Embryonic Stem ◽  
Stem Cell Differentiation ◽  
Embryonic Stem Cell Differentiation
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