Embryoid Body Differentiation of Mouse Embryonic Stem Cells into Neurectoderm and Neural Progenitors

Shparberg, Rachel; Glover, Hannah J; Morris, Mitchell

doi:10.1007/978-1-4939-9631-5_21

Cited by 12 publications

(7 citation statements)

References 28 publications

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“…Analogous to this, ESCs require medium supplementation with L-Gln to permit proliferation (Carey et al, 2015) and L-Pro to permit differentiation (Washington et al, 2010;Casalino et al, 2011;Shparberg et al, 2019a), even though both are produced endogenously. In the case of L-Pro, ESCs self-limit production by an autoregulatory loop mediated by free prolyl-tRNA (i.e., tRNA not loaded with proline) coupled to the aminoacid starvation response (AAR) pathway (D'Aniello et al, 2015).…”

Section: Discussionmentioning

confidence: 99%

Selected Amino Acids Promote Mouse Pre-implantation Embryo Development in a Growth Factor-Like Manner

Morris¹,

Ozsoy²,

Zada³

et al. 2020

Front. Physiol.

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Groups of amino acids, and some selected amino acids, added to media used for culture of pre-implantation embryos have previously been shown to improve development in various ways including survival to the blastocyst stage, increased blastocyst cell number and improved hatching. In this study, we cultured 1-cell mouse embryos for 5 days to the hatching blastocyst stage in isosmotic medium (270 mOsm/kg) at high density (10 embryos/10 µL), where autocrine/paracrine support of development occurs, and low density (1 embryo/100 µL), where autocrine/paracrine support is minimized and development is compromised. When 400 µM L-Pro or 1 mM L-Gln was added to embryos at low density, the percentage of embryos reaching the blastocyst stage and the percentage hatching increased compared to low-density culture without these amino acids, and were now similar to those for embryos cultured at high density without amino acids. When L-Pro or L-Gln was added to embryos at high density, the percentage of embryos reaching the blastocyst stage didn't change but hatching improved. Neither embryo culture density nor the presence of these amino acids had any effect on blastocyst cell number. D-Pro and the osmolytes Gly and Betaine did not improve embryo development in low-or high-density culture indicating the mechanism was stereospecific and not osmotic, respectively. L-Pro-and L-Glnmediated improvement in development is observed from the 5-cell stage and persists to the blastocyst stage. Molar excess of Gly, Betaine or L-Leu over L-Pro eliminated improvement in development and hatching consistent with them acting as competitive inhibitors of transporter-mediated uptake across the plasma membrane. The L-Pro effect is dependent on mTORC1 signaling (rapamycin sensitive) while that for L-Gln is not. The addition of L-Pro leads to significant nuclear translocation of p-Akt S473 at the 2-and 4-cell stages and of p-ERK1/2 T202/Y204 nuclear translocation at the 2-, 4-, and 8-cell stages. L-Pro improvement in embryo development involves mechanisms analogous to those seen with Pro-mediated differentiation of mouse ES cells, which is also stereoselective, dependent on transporter uptake, and activates Akt, ERK, and mTORC1 signaling pathways.

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Section: Discussionmentioning

confidence: 99%

Selected Amino Acids Promote Mouse Pre-implantation Embryo Development in a Growth Factor-Like Manner

Morris¹,

Ozsoy²,

Zada³

et al. 2020

Front. Physiol.

Self Cite

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“…This is one of the fastest and simplest methods for neural differentiation available since unlike other methods, we do not need to seed the generated EBs on adherent plates or harvest neural rosettes, which are additional complex steps often used. 50 A similar technique to ours has been employed recently by Hanafiah et al to obtain NPCs by Day 8 and neurons by Day 12, as demonstrated via the expression of nestin and neurofilament markers. 51 In our protocol, we obtain NPCs by Day 6 and supplement BDNF and βFGF in the neuronal media to ensure neuronal maturation over the following 3 weeks.…”

Section: Presence Of Erα-66 and Mers In Stem Cellsmentioning

confidence: 82%

“…With this protocol, we generated βIII‐tubulin positive cells by D7 (13 days total). This is one of the fastest and simplest methods for neural differentiation available since unlike other methods, we do not need to seed the generated EBs on adherent plates or harvest neural rosettes, which are additional complex steps often used 50 . A similar technique to ours has been employed recently by Hanafiah et al to obtain NPCs by Day 8 and neurons by Day 12, as demonstrated via the expression of nestin and neurofilament markers 51 .…”

Section: Discussionmentioning

confidence: 96%

Localisation of oestrogen receptors in stem cells and in stem cell‐derived neurons of the mouse

Davis

Vajaria

Delivopoulos

et al. 2022

J Neuroendocrinology

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Oestrogen receptors (ER) transduce the effects of the endogenous ligand, 17β-estradiol in cells to regulate a number of important processes such as reproduction, neuroprotection, learning and memory and anxiety. The ERα or ERβ are classical intracellular nuclear hormone receptors while some of their variants or novel proteins such as the G-protein coupled receptor (GPCR), GPER1/GPR30 are reported to localise in intracellular as well as plasma membrane locations. Although the brain is an

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“…Of note, embryonic stem (ES) cells are derived from blastomeres that form the inner cell mass of vertebrate embryos and are thought to retain this developmental pluripotent state. Lineages from all three embryonic layers, including germ cells, can be derived from ES cells (Ding et al, 2016;Shparberg et al, 2019;Vazin and Freed, 2010). Since there are no ES cell lines derived from C. elegans, this topic will not be addressed in this review, but we recommend (Huang et al, 2015) and (Eguizabal et al, 2019).…”

Section: Blastomeres and Progenitorsmentioning

confidence: 99%

On the origins and conceptual frameworks of natural plasticity—Lessons from single-cell models in C. elegans

Lambert

Lloret-Fernández

Laplane

et al. 2021

Current Topics in Developmental Biology

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How flexible are cell identities? This problem has fascinated developmental biologists for several centuries and can be traced back to Abraham Trembley's pioneering manipulations of Hydra to test its regeneration abilities in the 1700s. Since the cell theory in the mid-nineteenth century, developmental biology has been dominated by a single framework in which embryonic cells are committed to specific cell fates, progressively and irreversibly acquiring their differentiated identities. This hierarchical, unidirectional and irreversible view of cell identity has been challenged in the past decades through accumulative evidence that many cell types are more plastic than previously thought, even in intact organisms.The paradigm shift introduced by such plasticity calls into question several other key traditional concepts, such as how to define a differentiated cell or more generally cellular identity, and has brought new concepts, such as distinct cellular states. In this review, we want to contribute to this representation by attempting to clarify the conceptual and theoretical frameworks of cell plasticity and identity. In the context of these new frameworks we describe here an atlas of natural plasticity of the cell identity in C. elegans, including our current understanding of the cellular and molecular mechanisms at play. The worm further provides interesting cases at the borderlines of cellular plasticity that highlight the conceptual challenges still ahead. We then discuss a set of future questions and perspectives arising from the studies of natural plasticity in the worm, that are shared with other reprogramming and plasticity events across phyla. Part 1: IntroductionThe study of cellular plasticity is the study of changes in cellular identities. In this section we will discuss the notion of cellular identity in the animal kingdom with an emphasis on the nematode Caenorhabditis elegans. We will start by discussing how to consensually define it, the conceptual steps describing its acquisition during development and what cellular plasticity entails (Fig. 1A). We will place particular focus on cellular plasticity events involving the swap of differentiated identities. -How to define cellular identity?Historically, cell types have been identified on the basis of their morphology, location, function and where possible, cell lineage. In the last 40 years, the focus moved first towards using molecular markers, notably through in vivo visualisation using fluorescent fusion constructs (Chalfie et al., 1994;

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Embryoid Body Differentiation of Mouse Embryonic Stem Cells into Neurectoderm and Neural Progenitors

Cited by 12 publications

References 28 publications

Selected Amino Acids Promote Mouse Pre-implantation Embryo Development in a Growth Factor-Like Manner

Selected Amino Acids Promote Mouse Pre-implantation Embryo Development in a Growth Factor-Like Manner

Localisation of oestrogen receptors in stem cells and in stem cell‐derived neurons of the mouse

On the origins and conceptual frameworks of natural plasticity—Lessons from single-cell models in C. elegans

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