scholarly journals Consistent apparent Young’s modulus of human embryonic stem cells and derived cell types stabilized by substrate stiffness regulation promotes lineage specificity maintenance

2020 ◽  
Vol 9 (1) ◽  
Author(s):  
Anqi Guo ◽  
Bingjie Wang ◽  
Cheng Lyu ◽  
Wenjing Li ◽  
Yaozu Wu ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Fate ◽  
Cell Expansion ◽  
Embryonic Stem ◽  
Cell Types ◽  
Substrate Stiffness ◽  
Cell Stage ◽  
Cytoskeletal Organization ◽  
Lineage Specificity

Abstract Background Apparent Young’s modulus (AYM), which reflects the fundamental mechanical property of live cells measured by atomic force microscopy and is determined by substrate stiffness regulated cytoskeletal organization, has been investigated as potential indicators of cell fate in specific cell types. However, applying biophysical cues, such as modulating the substrate stiffness, to regulate AYM and thereby reflect and/or control stem cell lineage specificity for downstream applications, remains a primary challenge during in vitro stem cell expansion. Moreover, substrate stiffness could modulate cell heterogeneity in the single-cell stage and contribute to cell fate regulation, yet the indicative link between AYM and cell fate determination during in vitro dynamic cell expansion (from single-cell stage to multi-cell stage) has not been established. Results Here, we show that the AYM of cells changed dynamically during passaging and proliferation on substrates with different stiffness. Moreover, the same change in substrate stiffness caused different patterns of AYM change in epithelial and mesenchymal cell types. Embryonic stem cells and their derived progenitor cells exhibited distinguishing AYM changes in response to different substrate stiffness that had significant effects on their maintenance of pluripotency and/or lineage-specific characteristics. On substrates that were too rigid or too soft, fluctuations in AYM occurred during cell passaging and proliferation that led to a loss in lineage specificity. On a substrate with ‘optimal’ stiffness (i.e., 3.5 kPa), the AYM was maintained at a constant level that was consistent with the parental cells during passaging and proliferation and led to preservation of lineage specificity. The effects of substrate stiffness on AYM and downstream cell fate were correlated with intracellular cytoskeletal organization and nuclear/cytoplasmic localization of YAP. Conclusions In summary, this study suggests that optimal substrate stiffness regulated consistent AYM during passaging and proliferation reflects and contributes to hESCs and their derived progenitor cells lineage specificity maintenance, through the underlying mechanistic pathways of stiffness-induced cytoskeletal organization and the downstream YAP signaling. These findings highlighted the potential of AYM as an indicator to select suitable substrate stiffness for stem cell specificity maintenance during in vitro expansion for regenerative applications.

scholarly journals Pluripotency and Epigenetic Factors in Mouse Embryonic Stem Cell Fate Regulation

2015 ◽  
Vol 35 (16) ◽  
pp. 2716-2728 ◽  
Author(s):  
Lluis Morey ◽  
Alexandra Santanach ◽  
Luciano Di Croce
Keyword(s):  
Cell Fate ◽  
Cancer Progression ◽  
Molecular Mechanisms ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cell ◽  
Basic Research ◽  
Cell Types ◽  
Pluripotency Factors ◽  
Perfect System

Embryonic stem cells (ESCs) are characterized by their ability to self-renew and to differentiate into all cell types of a given organism. Understanding the molecular mechanisms that govern the ESC state is of great interest not only for basic research—for instance, ESCs represent a perfect system to study cellular differentiationin vitro—but also for their potential implications in human health, as these mechanisms are likewise involved in cancer progression and could be exploited in regenerative medicine. In this minireview, we focus on the latest insights into the molecular mechanisms mediated by the pluripotency factors as well as their roles during differentiation. We also discuss recent advances in understanding the function of the epigenetic regulators, Polycomb and MLL complexes, in ESC biology.

Human Embryonic Stem Cell-Derived Mesenchymal Stroma Cells (hES-MSC) Share Basic Properties with Bone Marrow MSC, They Have Potent Stroma Support Capacities but Lack Immune-Modulatory Function. Implications for off-the Shelf ES-MSC Therapies.

Blood ◽  
2010 ◽  
Vol 116 (21) ◽  
pp. 3858-3858 ◽  
Author(s):  
Ou Li ◽  
Ariane Tormin ◽  
Jan Claas Brune ◽  
Berit Sundberg ◽  
Johan Hyllner ◽  
...  
Keyword(s):  
Bone Marrow ◽  
Stem Cell ◽  
Human Embryonic Stem Cell ◽  
Embryonic Stem ◽  
Cell Types ◽  
Es Cell ◽  
Nsg Mice ◽  
Mesenchymal Stroma ◽  
Mesenchymal Stroma Cells

Abstract Abstract 3858 Mesenchymal stroma cells (MSC) have a high potential for novel cell therapy approaches in clinical transplantation due to their intriguing properties, e.g. high proliferation and differentiation capacity, stromal support and immune-modulation. Commonly, bone marrow-derived MSC (BM-MSC) are used for clinical MSC cell therapies. However, BM-derived MSC have a restricted proliferative capacity and cultured BM-MSC are heterogeneous and thus difficult to standardize. Human embryonic stem cell-derived mesenchymal stroma cells (hES-MSC) have recently been developed and might represent an alternative and unlimited source of hMSCs. We therefore aimed to characterize human ES-cell-derived MSC, i.e. the hES-MSC line hES-MP002.5 (Cellartis) and compare its properties with normal human bone marrow (BM) derived MSC. We found that hES-MP cells have lower yet reasonable CFU-F capacity when compared with BM-MSC (6+3 vs 25+1 CFU-F per 100 cells). hES-MP cells showed similar immunophenotypic properties compared with BM-MSC (flow cytometry): Both cell types were positive for CD105, CD73, CD166, HLA Class I, CD44, CD146 and CD90, and cells were negative for surface markers such as CD45, CD34, CD14, CD31, CD19, and HLA-DR. hES-MP, like BM-MSC, could be differentiated into adipocytes, osteoblasts and chondrocytes upon induction in vitro. In order to test whether MSC were capable of homing to the bone marrow after intravenous injection, hES-MP and BM-MSC were markerd with GFP, and sorted GFP-positive cells were injected intravenously into NOD.Cg-PrkdcscidIl2rgtm1Wjl/SzJ (NSG) mice. GFP-positive cells were not detected in the bone marrow 24 hours after injection, neither when hES-MP cells were injected, nor - and as expected - when cultured BM-MSC were used. Intra-femoral transplantation into NSG mice using GFP expressing hES-MP and BM-MSC on the other hand demonstrated successful long-term engraftment (8 weeks) for both cell types. Morphology and intra-femoral localization of hES-MP were similar compared to BM-MSC. LTC-IC and co-transplantation experiments with cord blood CD34+ hematopoietic cells demonstrated furthermore that hES-MP, like BM-MSC, possess potent stroma support function both in vitro and in vivo. However, hES-MP showed no or only little activity in mixed lymphocyte cultures and PHA lymphocyte stimulation assays. In summary, our data demonstrate that MSC derived from hES cells have biological properties and potent stroma functions similar to conventional BM-MSC. Thus, ES-cell derived MSC might be an attractive and reliable alternative and unlimited source for obtaining MSC for clinical cell therapy. However, hES-MP probably have no or only little immuno-modulative capacity, which may limit their potential clinical use. Disclosures: Hyllner: Cellartis AB: Employment.

scholarly journals Effects of Feeder Cell Types on Culture of Mouse Embryonic Stem Cell In Vitro

2015 ◽  
Vol 19 (3) ◽  
pp. 119-126 ◽  
Author(s):  
Yun-Gwi Park ◽  
Seung-Eun Lee ◽  
Eun-Young Kim ◽  
Hyuk Hyun ◽  
Min-Young Shin ◽  
...  
Keyword(s):  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Embryonic Stem ◽  
Mouse Embryonic Stem Cell ◽  
Cell Types ◽  
Feeder Cell

scholarly journals Dlk1-Dio3 locus-derived lncRNAs perpetuate postmitotic motor neuron cell fate and subtype identity

eLife ◽  
2018 ◽  
Vol 7 ◽  
Author(s):  
Ya-Ping Yen ◽  
Wen-Fu Hsieh ◽  
Ya-Yin Tsai ◽  
Ya-Lin Lu ◽  
Ee Shan Liau ◽  
...  
Keyword(s):  
Stem Cell ◽  
Cell Fate ◽  
Motor Neurons ◽  
Neural Development ◽  
Hox Genes ◽  
Critical Role ◽  
Embryonic Stem ◽  
Cell Types ◽  
Aberrant Expression ◽  
Spinal Cord Development

The mammalian imprinted Dlk1-Dio3 locus produces multiple long non-coding RNAs (lncRNAs) from the maternally inherited allele, including Meg3 (i.e., Gtl2) in the mammalian genome. Although this locus has well-characterized functions in stem cell and tumor contexts, its role during neural development is unknown. By profiling cell types at each stage of embryonic stem cell-derived motor neurons (ESC~MNs) that recapitulate spinal cord development, we uncovered that lncRNAs expressed from the Dlk1-Dio3 locus are predominantly and gradually enriched in rostral motor neurons (MNs). Mechanistically, Meg3 and other Dlk1-Dio3 locus-derived lncRNAs facilitate Ezh2/Jarid2 interactions. Loss of these lncRNAs compromises the H3K27me3 landscape, leading to aberrant expression of progenitor and caudal Hox genes in postmitotic MNs. Our data thus illustrate that these lncRNAs in the Dlk1-Dio3 locus, particularly Meg3, play a critical role in maintaining postmitotic MN cell fate by repressing progenitor genes and they shape MN subtype identity by regulating Hox genes.

scholarly journals Controlling Self-Renewal and Differentiation of Stem Cells via Mechanical Cues

2012 ◽  
Vol 2012 ◽  
pp. 1-12 ◽  
Author(s):  
Michele M. Nava ◽  
Manuela T. Raimondi ◽  
Riccardo Pietrabissa
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Cell Fate ◽  
Cell Response ◽  
Recent Literature ◽  
Focal Adhesions ◽  
Substrate Stiffness ◽  
Self Renewal ◽  
Mechanical Factors

The control of stem cell responsein vitro, including self-renewal and lineage commitment, has been proved to be directed by mechanical cues, even in the absence of biochemical stimuli. Through integrin-mediated focal adhesions, cells are able to anchor onto the underlying substrate, sense the surrounding microenvironment, and react to its properties. Substrate-cell and cell-cell interactions activate specific mechanotransduction pathways that regulate stem cell fate. Mechanical factors, including substrate stiffness, surface nanotopography, microgeometry, and extracellular forces can all have significant influence on regulating stem cell activities. In this paper, we review all the most recent literature on the effect of purely mechanical cues on stem cell response, and we introduce the concept of “force isotropy” relevant to cytoskeletal forces and relevant to extracellular loads acting on cells, to provide an interpretation of how the effects of insoluble biophysical signals can be used to direct stem cells fatein vitro.

scholarly journals Developing defined substrates for stem cell culture and differentiation

2018 ◽  
Vol 373 (1750) ◽  
pp. 20170230 ◽  
Author(s):  
Louise Hagbard ◽  
Katherine Cameron ◽  
Paul August ◽  
Christopher Penton ◽  
Malin Parmar ◽  
...  
Keyword(s):  
Extracellular Matrix ◽  
Stem Cell ◽  
Cell Fate ◽  
Cell Types ◽  
Specific Cell ◽  
Biologically Relevant ◽  
Stem Cell Maintenance ◽  
Signalling Molecules ◽  
Cell Matrix

Over the past few decades, a variety of different reagents for stem cell maintenance and differentiation have been commercialized. These reagents share a common goal in facilitating the manufacture of products suitable for cell therapy while reducing the amount of non-defined components. Lessons from developmental biology have identified signalling molecules that can guide the differentiation process in vitro , but less attention has been paid to the extracellular matrix used. With the introduction of more biologically relevant and defined matrices, that better mimic specific cell niches, researchers now have powerful resources to fine-tune their in vitro differentiation systems, which may allow the manufacture of therapeutically relevant cell types. In this review article, we revisit the basics of the extracellular matrix, and explore the important role of the cell–matrix interaction. We focus on laminin proteins because they help to maintain pluripotency and drive cell fate specification. This article is part of the theme issue ‘Designer human tissue: coming to a lab near you’.

scholarly journals An in vitro stem cell model of human epiblast and yolk sac interaction

eLife ◽  
2021 ◽  
Vol 10 ◽  
Author(s):  
Kirsty ML Mackinlay ◽  
Bailey AT Weatherbee ◽  
Viviane Souza Rosa ◽  
Charlotte E Handford ◽  
George Hudson ◽  
...  
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Cell Fate ◽  
Human Embryo ◽  
Cell Model ◽  
Embryonic Stem ◽  
Yolk Sac ◽  
Embryonic Cells ◽  
Post Implantation

Human embryogenesis entails complex signalling interactions between embryonic and extra-embryonic cells. However, how extra-embryonic cells direct morphogenesis within the human embryo remains largely unknown due to a lack of relevant stem cell models. Here, we have established conditions to differentiate human pluripotent stem cells (hPSCs) into yolk sac-like cells (YSLCs) that resemble the post-implantation human hypoblast molecularly and functionally. YSLCs induce the expression of pluripotency and anterior ectoderm markers in human embryonic stem cells (hESCs) at the expense of mesoderm and endoderm markers. This activity is mediated by the release of BMP and WNT signalling pathway inhibitors, and, therefore, resembles the functioning of the anterior visceral endoderm signalling centre of the mouse embryo, which establishes the anterior-posterior axis. Our results implicate the yolk sac in epiblast cell fate specification in the human embryo and propose YSLCs as a tool for studying post-implantation human embryo development in vitro.

scholarly journals Amniotic Fluid and Amniotic Membrane Stem Cells: Marker Discovery

2012 ◽  
Vol 2012 ◽  
pp. 1-9 ◽  
Author(s):  
Maria G. Roubelakis ◽  
Ourania Trohatou ◽  
Nicholas P. Anagnou
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Amniotic Fluid ◽  
Amniotic Membrane ◽  
Adult Stem Cells ◽  
Embryonic Stem ◽  
Cell Types ◽  
Stem Cell Population ◽  
Marker Discovery

Amniotic fluid (AF) and amniotic membrane (AM) have been recently characterized as promising sources of stem or progenitor cells. Both not only contain subpopulations with stem cell characteristics resembling to adult stem cells, such as mesenchymal stem cells, but also exhibit some embryonic stem cell properties like (i) expression of pluripotency markers, (ii) high expansion in vitro, or (iii) multilineage differentiation capacity. Recent efforts have been focused on the isolation and the detailed characterization of these stem cell types. However, variations in their phenotype, their heterogeneity described by different groups, and the absence of a single marker expressed only in these cells may prevent the isolation of a pure homogeneous stem cell population from these sources and their potential use of these cells in therapeutic applications. In this paper, we aim to summarize the recent progress in marker discovery for stem cells derived from fetal sources such as AF and AM, using novel methodologies based on transcriptomics, proteomics, or secretome analyses.

scholarly journals Potential Clinical Applications for Human Pluripotent Stem Cell-Derived Blood Components

2011 ◽  
Vol 2011 ◽  
pp. 1-11 ◽  
Author(s):  
Erin A. Kimbrel ◽  
Shi-Jiang Lu
Keyword(s):  
Stem Cells ◽  
Stem Cell ◽  
Large Scale ◽  
Embryonic Stem ◽  
Adult Stem Cell ◽  
Cell Types ◽  
The Body ◽  
Blood Components ◽  
Induced Pluripotent

The ability of human embryonic stem cells (hESCs) and induced pluripotent stem cells (iPSCs) to divide indefinitely without losing pluripotency and to theoretically differentiate into any cell type in the body makes them highly attractive cell sources for large scale regenerative medicine purposes. The current use of adult stem cell-derived products in hematologic intervention sets an important precedent and provides a guide for developing hESC/iPSC based therapies for the blood system. In this review, we highlight biological functions of mature cells of the blood, clinical conditions requiring the transfusion or stimulation of these cells, and the potential for hESC/iPSC-derivatives to serve as functional replacements. Many researchers have already been able to differentiate hESCs and/or iPSCs into specific mature blood cell types. For example, hESC-derived red blood cells and platelets are functional in tasks such as oxygen delivery and blood clotting, respectively and may be able to serve as substitutes for their donor-derived counterparts in emergencies. hESC-derived dendritic cells are functional in antigen-presentation and may be used as off-the-shelf vaccine therapies to stimulate antigen-specific immune responses against cancer cells. However,in vitrodifferentiation systems used to generate these cells will need further optimization before hESC/iPSC-derived blood components can be used clinically.

scholarly journals Hydrogel platform for in vitro three-dimensional assembly of human stem cell-derived β cells and endothelial cells

2019 ◽  
Author(s):  
Punn Augsornworawat ◽  
Leonardo Velazco-Cruz ◽  
Jiwon Song ◽  
Jeffrey R. Millman
Keyword(s):  
Endothelial Cells ◽  
Stem Cell ◽  
Embryonic Stem Cell ◽  
Human Embryonic Stem Cell ◽  
Three Dimensional ◽  
Embryonic Stem ◽  
Cell Types ◽  
Diabetic Patients ◽  
Β Cells

AbstractDifferentiation of stem cells into functional replacement cells and tissues is a major goal of the regenerative medicine field. However, one limitation has been organization of differentiated cells into multi-cellular, three-dimensional assemblies. The islets of Langerhans contain many endocrine and non-endocrine cell types, such as insulin-producing β cells and endothelial cells. Transplantation of exogenous islets into diabetic patients can serve as a cell replacement therapy, replacing the need for patients to inject themselves with insulin, but the number of available islets from cadaveric donors is low. We have developed a strategy of assembling human embryonic stem cell-derived β cells with endothelial cells into three-dimensional aggregates on a hydrogel. The resulting islet organoids express β cell markers and are functional, capable of undergoing glucose-stimulated insulin secretion. These results provide a platform for evaluating the effects of the islet tissue microenvironment on human embryonic stem cell-derived β cells and other islet endocrine cells to develop tissue engineered islets.

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