Hydrophobic surfaces for enhanced differentiation of embryonic stem cell-derived embryoid bodies

Department of Molecular and Medical Pharmacology, David Geffen School of Medicine, University of California, Los Angeles, CA 90095, USA.
Proceedings of the National Academy of Sciences (Impact Factor: 9.67). 10/2008; 105(38):14459-64. DOI: 10.1073/pnas.0807235105
Source: PubMed


With their unique ability to differentiate into all cell types, embryonic stem (ES) cells hold great therapeutic promise. To improve the efficiency of embryoid body (EB)-mediated ES cell differentiation, we studied murine EBs on the basis of their size and found that EBs with an intermediate size (diameter 100-300 microm) are the most proliferative, hold the greatest differentiation potential, and have the lowest rate of cell death. In an attempt to promote the formation of this subpopulation, we surveyed several biocompatible substrates with different surface chemical parameters and identified a strong correlation between hydrophobicity and EB development. Using self-assembled monolayers of various lengths of alkanethiolates on gold substrates, we directly tested this correlation and found that surfaces that exhibit increasing hydrophobicity enrich for the intermediate-size EBs. When this approach was applied to the human ES cell system, similar phenomena were observed. Our data demonstrate that hydrophobic surfaces serve as a platform to deliver uniform EB populations and may significantly improve the efficiency of ES cell differentiation.

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Available from: Eric Gschweng, Oct 07, 2015
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    • "The differences in the rate of diffusion of biologically active substances in different cell layers of EBs probably determine different direction and effectiveness of their differentiation into different types of embryonic cells (Bauwens et al., 2008; Messana et al., 2008; Mohr et al., 2010; Valamehr et al., 2008). The threeedimensionality of EBs, as well as embryos, per se provides a concentraa tion gradient of substances that promotes the developp ment and protection of cells from damaging factors. "
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    ABSTRACT: The initial stages of in vitro differentiation of embryonic stem cells are considered as unique three-dimensional models of early development of mammals for basic, pharmacological, and toxicological studies. It has been previously shown (Gordeeva, 2012) that the assessment of embryotoxicity in the model of undifferentiated embryonic stem cells can be insufficiently accurate in predicting toxic effects on mammalian embryos. In view of this, we performed a comparative study of the damaging effects of the cytostatic etoposide in undifferentiated embryonic stem cells and embryoid bodies of different stages of differentiation that have similar three-dimensional structures with early embryos. The analysis of growth, cell death, and dynamics of differentiation of embryonic stem cells and embryoid bodies exposed to etoposide showed that the cytostatic and cytotoxic effects of etoposide are stage-specific. The damaging effects of etoposide were maximum in the undifferentiated embryonic stem cells and decreased with growth and differentiation of embryoid bodies. We suggest that the increase of embryoid body volume and overgrowth of extraembryonic endoderm layer lead to a decrease in the diffusion, transport and metabolism of chemical and bioactive substances and prevent the damaging effects.
    Russian Journal of Developmental Biology 11/2013; 44(6):283-289. DOI:10.1134/S1062360413060039 · 0.31 Impact Factor
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    • "Toworfe and his coworkers reported that fibronectin-coated PDMS could enhance and upgrade MC3T3-E1 cellular functions, particularly on its attachment of and spreading on the PDMS surfaces12. Many other studies have also proven that PDMS surfaces, as well as cellular microenvironment, could affect and regulate embryonic and functional stem cell fates131415. The PDMS topography and stiffness also have micro-environmental effects on the differentiation of human epidermal stem cells, mesenchymal stem cells, and others131415. "
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    ABSTRACT: This systematic study clarified a few interfacial aspects of cancer cell phenotypes on polydimethylsiloxane (PDMS) substrates and indicated that the cell phenotypic equilibrium greatly responds to cell-to-surface interactions. We demonstrated that coatings of fibronectin, bovine serum albumin (BSA), or collagen with or without oxygen-plasma treatments of the PDMS surfaces dramatically impacted the phenotypic equilibrium of breast cancer stem cells, while the variations of the PDMS elastic stiffness had much less such effects. Our results showed that the surface coatings of collagen and fibronectin on PDMS maintained breast cancer cell phenotypes to be nearly identical to the cultures on commercial polystyrene Petri dishes. The surface coating of BSA provided a weak cell-substrate adhesion that stimulated the increase in stem-cell-like subpopulation. Our observations may potentially guide surface modification approaches to obtain specific cell phenotypes.
    Scientific Reports 07/2013; 3:2332. DOI:10.1038/srep02332 · 5.58 Impact Factor
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    • "Several studies have shown that heterogeneity in human ESC colony size and resulting EB aggregate size results in variability in differentiation experiments and significant decreases in differentiation yields [23–26]. The effect of EB homogeneity on differentiation potential of human iPSCs into cardiomyocytes was examined. "
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    ABSTRACT: Proper maintenance of stem cells is essential for successful utilization of ESCs/iPSCs as tools in developmental and drug discovery studies and in regenerative medicine. Standardization is critical for all future applications of stem cells and necessary to fully understand their potential. This study reports a novel approach for the efficient, consistent expansion of human ESCs and iPSCs using laser sectioning, instead of mechanical devices or enzymes, to divide cultures into defined size clumps for propagation. Laser-mediated propagation maintained the pluripotency, quality, and genetic stability of ESCs/iPSCs and led to enhanced differentiation potential. This approach removes the variability associated with ESC/iPSC propagation, significantly reduces the expertise, labor, and time associated with manual passaging techniques and provides the basis for scalable delivery of standardized ESC/iPSC lines. Adoption of standardized protocols would allow researchers to understand the role of genetics, environment, and/or procedural effects on stem cells and would ensure reproducible production of stem cell cultures for use in clinical/therapeutic applications.
    05/2012; 2012:926463. DOI:10.1155/2012/926463
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