Activin A Suppresses Induced Formation of Neuroectoderm in Colonies of Parthenogenetic Stem Cells In Vitro

2011 ◽  
Vol 151 (4) ◽  
pp. 502-505 ◽  
Author(s):  
R. A. Semechkin ◽  
T. V. Abramihina ◽  
D. A. Isaev
Keyword(s):  
Stem Cells ◽  
Activin A

The effect of dual inhibition of Ras–MEK–ERK and GSK3β pathways on development of in vitro cultured rabbit embryos

Zygote ◽  
2020 ◽  
Vol 28 (3) ◽  
pp. 183-190 ◽  
Author(s):  
Babett Bontovics ◽  
Pouneh Maraghechi ◽  
Bence Lázár ◽  
Mahek Anand ◽  
Kinga Németh ◽  
...  
Keyword(s):  
Stem Cells ◽  
Activin A ◽  
Mouse Strains ◽  
Suitable Model ◽  
Embryonic Cells ◽  
Cell Stage ◽  
Human Ipscs ◽  
Dual Inhibition ◽  
Rabbit Embryos

SummaryDual inhibition (2i) of Ras–MEK–ERK and GSK3β pathways enables the derivation of embryo stem cells (ESCs) from refractory mouse strains and, for permissive strains, allows ESC derivation with no external protein factor stimuli involvement. In addition, blocking of ERK signalling in 8-cell-stage mouse embryos leads to ablation of GATA4/6 expression in hypoblasts, suggesting fibroblast growth factor (FGF) dependence of hypoblast formation in the mouse. In human, bovine or porcine embryos, the hypoblast remains unaffected or displays slight-to-moderate reduction in cell number. In this study, we demonstrated that segregation of the hypoblast and the epiblast in rabbit embryos is FGF independent and 2i treatment elicits only a limited reinforcement in favour of OCT4-positive epiblast populations against the GATA4-/6-positive hypoblast population. It has been previously shown that TGFβ/Activin A inhibition overcomes the pervasive differentiation and inhomogeneity of rat iPSCs, rat ESCs and human iPSCs while prompting them to acquire naïve properties. However, TGFβ/Activin A inhibition, alone or together with Rho-associated, coiled-coil containing protein kinase (ROCK) inhibition, was not compatible with the viability of rabbit embryos according to the ultrastructural analysis of preimplantation rabbit embryos by electron microscopy. In rabbit models ovulation upon mating allows the precise timing of progression of the pregnancy. It produces several embryos of the desired stage in one pregnancy and a relatively short gestation period, making the rabbit embryo a suitable model to discover the cellular functions and mechanisms of maintenance of pluripotency in embryonic cells and the embryo-derived stem cells of other mammals.

scholarly journals The crucial role of Activin A on the formation of primordial germ cell-like cells from skin-derived stem cells in vitro

Cell Cycle ◽  
2015 ◽  
Vol 14 (19) ◽  
pp. 3016-3029 ◽  
Author(s):  
Rui Sun ◽  
Yuan-Chao Sun ◽  
Wei Ge ◽  
Hui Tan ◽  
Shun-Feng Cheng ◽  
...  
Keyword(s):  
Stem Cells ◽  
Germ Cell ◽  
Crucial Role ◽  
Primordial Germ Cell ◽  
Activin A

scholarly journals Different Phenotypes and Chondrogenic Responses of Human Menstrual Blood and Bone Marrow Mesenchymal Stem Cells to activin A and TGF-β3

2020 ◽  
Author(s):  
Ilona Uzieliene ◽  
Edvardas Bagdonas ◽  
Kazuto Hoshi ◽  
Tomoaki Sakamoto ◽  
Atsuhiko Hikita ◽  
...  
Keyword(s):  
Stem Cells ◽  
Mesenchymal Stem Cells ◽  
Chondrogenic Differentiation ◽  
Collagen Type ◽  
Activin A ◽  
Cell Surface Markers ◽  
Type Ii ◽  
Marrow Mesenchymal Stem Cells

Abstract Background: Due to its low capacity for self-repair, articular cartilage is highly susceptible to damage and deterioration, which leads to the development of degenerative joint diseases such as osteoarthritis. Menstrual blood-derived mesenchymal stem cells (MenSCs) are much less characterized compared to bone marrow mesenchymal stem cells (BMMSCs). However, MenSCs seem an attractive alternative to classical BMMSCs due to ease of access and broader differentiation capacity. The aim of this study was to evaluate chondrogenic differentiation potential of MenSCs and BMMSCs stimulated with transforming growth factor β (TGF-β3) and activin A, member of the TGF-β superfamily of proteins.Methods: MenSCs (n=6) and BMMSCs (n=5) were isolated from different healthy donors. Expression of cell surface markers CD90, CD73, CD105, CD44, CD45, CD14, CD36, CD55, CD54, CD63, CD106, CD34, CD10, Notch1 was analysed by flow cytometry. Cell proliferation capacity was determined using CCK-8 proliferation kit. Adipogenic differentiation capacity was evaluated according to Oil-Red staining, osteogenic differentiation - Alizarin Red staining. Chondrogenic differentiation (Activin A and TGF-β3 stimulation) was induced in vitro and in vivo (subcutaneous scaffolds in nude BALB/c mice) and investigated by histologically and by expression of chondrogenic genes (collagen type II, aggrecan). Activin A protein production was evaluated by ELISA.Results: MenSCs exhibited a higher proliferation rate, as compared to BMMSCs, and a different expression profile of several cell surface markers. Activin A stimulated collagen type II gene expression and glycosaminoglycan synthesis in TGF-β3 treated MenSCs but not in BMMSCs, both in vitro and in vivo, although the effects of TGF-β3 alone were more pronounced in BMMSCs in vitro. Conclusion: These data suggest that activin A exerts differential effects on the induction of chondrogenic differentiation in MenSCs vs. BMMSCs, which implies that different mechanisms of chondrogenic regulation are activated in these cells. Following further optimisation of differentiation protocols and the choice of growth factors, potentially including activin A, MenSCs may turn out to be a promising population of stem cells for the development of cell-based therapies with the capacity to stimulate cartilage repair and regeneration.Trial registration: Not applicable.

Human CD34+ Haematopoietic Stem Cells (HSC) from Umbilical Cord Blood Display an Endodermal Phenotype When Exposed to Activin A In Vitro.

Blood ◽  
2005 ◽  
Vol 106 (11) ◽  
pp. 1697-1697
Author(s):  
Claudia Albera ◽  
Anne E. Bishop ◽  
Julia M. Polak ◽  
Nicki Panoskaltsis
Keyword(s):  
Stem Cells ◽  
Cord Blood ◽  
Umbilical Cord ◽  
Umbilical Cord Blood ◽  
Activin A ◽  
Rt Pcr ◽  
Hematopoietic Stem ◽  
Transcript Levels ◽  
Time Points

Abstract Adult stem cells may not be tissue specific and transdifferentiation into cells committed to other tissues may occur. In vivo, bone marrow HSC can contribute to cells characteristic of resident endoderm-derived tissues, such as lung. We hypothesized that if transdifferentiation can occur, then in vitro evidence could be obtained from an enriched population of hematopoietic cells. This study evaluated whether endodermal lineages could be derived from human umbilical cord blood (UCB) cells in order to provide a novel source for the regeneration of non-hematopoietic tissues, such as the lung. UCB CD34+ hematopoietic stem cells were magnetically enriched to a purity greater than 92%. A fraction of these was immediately characterised by flow cytometry and the remaining cells were expanded for 3 days in serum-free media. The effects of a combination of growth factors on these cells, including FGF-4, HGF and Activin A, were evaluated after 15 and 30 days of culture. RT-PCR analysis revealed detectable mRNA levels from representative endodermal genes (SOX17, GATA4 and FOXa2) after 15 days of induction and epithelial genes cytokeratin (CK)-18 and E-cadherin (E-cad) after 30 days of induction. Control cells after 3 days expansion and freshly isolated CD34+ (day 0) showed detectable transcript levels of HOXb4, Gata1 and PECAM-1 but they were negative for all endoderm markers. A panel of endodermal and epithelial primary antibodies, GATA4, GATA6, E-cad, CK-18, Pan-CK and Albumin were immunolocalized in cultures after 15 and 30 days of induction showing gene and protein expression. In order to evaluate the temporal expression pattern of the endodermal genes excluding the endothelial cell component, enriched CD34+ cells were subsequently FACS-sorted to yield a CD34+CD45+ cell population of greater than 98% purity. Quantitative real time RT-PCR was used to measure fold changes in the expression of GATA4, FOXa2, SOX17, E-cad and CK-18 genes in response to low and high Activin A concentrations at 6 different time points of induction cell culture (0, 3, 5, 7, 9, 12 and 15 days). Exposure to Activin A increased the expression of the endoderm transcript levels as early as 3 days of induction for E-cad and SOX17 and at 7 and 9 days of induction for GATA4 and FOXa2, respectively. Data were normalized to the housekeeping gene ß-actin and were expressed relative to the level of the gene of interest at time zero. HSC cultivated without Activin A and evaluated at the same time points did not express endoderm transcripts. These findings show that in the presence of Activin A, hematopoietic stem cells can be induced to express genes and proteins indicative of endoderm lineage in vitro. Since Activin A is upregulated in damaged tissue and is required for wound healing, the data suggest a mechanism for in vitro “transdifferentiation” of HSC, especially in the repair of damaged organs, and indicate that this process may be applied therapeutically for regenerative medicine.

scholarly journals Tumorigenic and Differentiation Potentials of Embryonic Stem Cells Depend on TGFβFamily Signaling: Lessons from Teratocarcinoma Cells Stimulated to Differentiate with Retinoic Acid

2017 ◽  
Vol 2017 ◽  
pp. 1-14 ◽  
Author(s):  
Olga Gordeeva ◽  
Sergey Khaydukov
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Retinoic Acid ◽  
Es Cells ◽  
Embryonic Stem ◽  
Activin A ◽  
Bmp Signaling ◽  
Induced Differentiation ◽  
Ec Cells

A significant challenge for the development of safe pluripotent stem cell-based therapies is the incomplete in vitro differentiation of the pluripotent stem cells and the presence of residual undifferentiated cells initiating teratoma development after transplantation in recipients. To understand the mechanisms of incomplete differentiation, a comparative study of retinoic acid-induced differentiation of mouse embryonic stem (ES) and teratocarcinoma (EC) cells was conducted. The present study identified differences in proliferative activity, differentiation, and tumorigenic potentials between ES and EC cells. Higher expression of Nanog and Mvh, as well as Activin A and BMP4, was found in undifferentiated ES cells than in EC cells. However, the expression levels of Activin A and BMP4 increased more sharply in the EC cells during retinoic acid-induced differentiation. Stimulation of the Activin/Nodal and BMP signaling cascades and inhibition of the MEK/ERK and PI3K/Act signaling pathways resulted in a significant decrease in the number of Oct4-expressing ES cells and a loss of tumorigenicity, similar to retinoic acid-stimulated EC cells. Thus, this study demonstrates that a differentiation strategy that modulates prodifferentiation and antiproliferative signaling in ES cells may be effective for eliminating tumorigenic cells and may represent a valuable tool for the development of safe stem cell therapeutics.

scholarly journals Thyrotropin-Independent Induction of Thyroid Endoderm from Embryonic Stem Cells by Activin A

Endocrinology ◽  
2008 ◽  
Vol 150 (4) ◽  
pp. 1970-1975 ◽  
Author(s):  
Risheng Ma ◽  
Rauf Latif ◽  
Terry F. Davies
Keyword(s):  
Stem Cells ◽  
Embryonic Stem Cells ◽  
Embryonic Stem ◽  
Activin A ◽  
Embryoid Bodies ◽  
Sodium Iodide Symporter ◽  
Thyroid Cells ◽  
Gene And Protein Expression ◽  
Igf I

To model the differentiation of thyroid epithelial cells, we examined embryoid bodies derived from undifferentiated murine embryonic stem cells treated with activin A to induce endoderm differentiation, the germ layer from which thyroid cells occur. The resulting endodermal cells were then further exposed to TSH and/or IGF-I for up to 21 d. Oct-4 and REX1 expression, required to sustain stem cell self-renewal and pluripotency, were appropriately down-regulated, whereas GATA-4, and α-fetoprotein, both endodermal-specific markers, increased as the embryonic stem cells were exposed to activin A. By d 5 culture, TSH receptor (TSHR) and sodium iodide symporter (NIS) gene and protein expression were markedly induced. Cells isolated by the fluorescence-activated cell sorter simultaneously expressed not only TSHR and NIS proteins but also PAX8 mRNA, an expression pattern unique to thyroid cells and expected in committed thyroid progenitor cells. Such expression continued until d 21 with no influence seen by the addition of TSH or IGF-I. The sequence of gene expression changes observed in these experiments demonstrated the emergence of definitive thyroid endoderm. The activin A induction of thyroid-specific markers, NIS and TSHR, occurred in the absence of TSH stimulation, and, therefore, the emergence of thyroid endoderm in vitro paralleled the emergence of thyroid cells in TSHR-knockout mice. Activin A is clearly a major regulator of thyroid endoderm.

scholarly journals Differential response of epiblast stem cells to Nodal and Activin signalling: a paradigm of early endoderm development in the embryo

2014 ◽  
Vol 369 (1657) ◽  
pp. 20130550 ◽  
Author(s):  
Keren Kaufman-Francis ◽  
Hwee Ngee Goh ◽  
Yoji Kojima ◽  
Joshua B. Studdert ◽  
Vanessa Jones ◽  
...  
Keyword(s):  
Stem Cells ◽  
Transforming Growth Factor ◽  
Activin A ◽  
Embryoid Bodies ◽  
Similar Process ◽  
Endoderm Differentiation ◽  
Epiblast Stem Cells ◽  
Suboptimal Outcome ◽  
Endoderm Development

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.

scholarly journals Activin A Modulates CRIPTO-1/HNF4α+ Cells to Guide Cardiac Differentiation from Human Embryonic Stem Cells

2017 ◽  
Vol 2017 ◽  
pp. 1-17 ◽  
Author(s):  
Robin Duelen ◽  
Guillaume Gilbert ◽  
Abdulsamie Patel ◽  
Nathalie de Schaetzen ◽  
Liesbeth De Waele ◽  
...  
Keyword(s):  
Stem Cells ◽  
Cell Fate ◽  
Heart Development ◽  
Embryoid Body ◽  
Embryonic Stem ◽  
Activin A ◽  
Embryoid Bodies ◽  
Cardiac Differentiation ◽  
Potential Applications

The use of human pluripotent stem cells in basic and translational cardiac research requires efficient differentiation protocols towards cardiomyocytes. In vitro differentiation yields heterogeneous populations of ventricular-, atrial-, and nodal-like cells hindering their potential applications in regenerative therapies. We described the effect of the growth factor Activin A during early human embryonic stem cell fate determination in cardiac differentiation. Addition of high levels of Activin A during embryoid body cardiac differentiation augmented the generation of endoderm derivatives, which in turn promoted cardiomyocyte differentiation. Moreover, a dose-dependent increase in the coreceptor expression of the TGF-β superfamily member CRIPTO-1 was observed in response to Activin A. We hypothesized that interactions between cells derived from meso- and endodermal lineages in embryoid bodies contributed to improved cell maturation in early stages of cardiac differentiation, improving the beating frequency and the percentage of contracting embryoid bodies. Activin A did not seem to affect the properties of cardiomyocytes at later stages of differentiation, measuring action potentials, and intracellular Ca2+ dynamics. These findings are relevant for improving our understanding on human heart development, and the proposed protocol could be further explored to obtain cardiomyocytes with functional phenotypes, similar to those observed in adult cardiac myocytes.

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