Formation of embryoid bodies from mouse embryonic stem cells cultured on silicon-coated surfaces

Fathi, Fardin; Altiraihi, T; Mowla, Seyed Javad; Movahedin, Mansoureh

doi:10.1007/s10616-009-9188-x

Cited by 6 publications

(4 citation statements)

References 30 publications

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“…Immediately before stimulation, tubes were coated with Sigmacote ® (Sigma-Aldrich), rinsed with distilled water, and dried with pressurised air in order to form a hydrophobic surface. The use of coated-tubes avoids attachment of membrane fragments components to the tube surface (19) and allows recovering larger concentrations of MPs/μl compared to non-coated tubes. Suspension of isolated platelet MPs (PMPs) was prepared as follows.…”

Section: Platelet Isolation and Stimulationmentioning

confidence: 99%

An innovative flow cytometric approach for small-size platelet microparticles: Influence of calcium

Montoro-García

Shantsila²,

Orenes‐Piñero³

et al. 2012

Thromb Haemost

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SummaryMicroparticles (MPs) are small submicron membrane-derived vesicles shed from a variety of cells and they have been implicated in different disorders. Accordingly, understanding of physiological characteristics of MPs and improvement of methods of their quantification are important for further advance in the field. Although flow cytometry is the most widely applied technique for MP analysis, it is limited by lack of adequate standardisation. Annexin V (AnV), which binds surface phos-phatidylserine (PS) with high affinity, has been long regarded as a marker of MPs, but AnV binding is Ca2+-dependent and it is unclear how [Ca2+] concentrations could affect AnV binding to MPs and its enumeration. MPs from citrated and heparinised plasma were labelled with AnV, anti-CD42b and quantified using an Apogee A50 flow cytometer. The small-size MP gate was defined with the use of size beads (from 0.1 to 0.5 μm) and confirmed with an in vitro assessment of platelet stimulation. Biotinylated anti-CD42b antibodies were then bound to streptavidin conjugated with different fluorochromes, leading to an amplified signal of platelet MPs (PMPs). Moderate increase of [Ca2+] concentrations in the annexin V staining buffer allows initial plasma recalcification and more accurate MP quantification in citrated plasma. Thrombin stimulation of platelet-free plasma containing only MPs did not produce any changes in the concentration of AnV+ MPs, but decreased the anti-CD42b binding. The results also indicate that prolonged storage and thrombin induce the release of AnV+ MPs whereas PS exposure in pre-existent MPs is not affected by thrombin. In conclusion, we present a sensitive protocol for the analysis of circulating and in vitro induced small-size PMPs that might contribute to future cardiovascular and clinical research.Note: The editorial process for this article was fully handled by Prof. Christian Weber, Editor-in-Chief.

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Section: Platelet Isolation and Stimulationmentioning

confidence: 99%

An innovative flow cytometric approach for small-size platelet microparticles: Influence of calcium

Montoro-García

Shantsila²,

Orenes‐Piñero³

et al. 2012

Thromb Haemost

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“…[37] Since hydrophobicity has been shown to play a role in bacterial attachment to surfaces, [38] we tested different silanes as coatings for long term biofilm culture on glass coverslips to reduce the surface zeta potential and increase the interaction strength with the bacterial surfaces. We selected (3-aminopropyl) triethoxysilane (APTES) and Sigmacote, two silanes extensively used as a surface coating to support stem cell cultures [39,40] in order to achieve stronger, long term adhesion due to the electrostatic interaction between the bacteria and the treated glass surface.…”

Section: Characterization Of Living Biointerfacesmentioning

confidence: 99%

Living Biointerfaces for the Maintenance of Mesenchymal Stem Cell Phenotypes

Petaroudi

Rodrigo‐Navarro

Dobre

et al. 2022

Adv Funct Materials

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Living interfaces are established as a novel class of active materials that aim to provide an alternative to traditional static cell culture methods by enabling users to accurately control cell behaviour in a precise, dynamic, and reliable system‐internal manner. To this day, the only reported biointerface has been a coculture between a biofilm of nonpathogenic genetically engineered bacteria and mammalian cells, where the recombinant proteins produced by the bacteria directly influence cell behaviour. In this work, a biointerface is presented between Lactococcus lactis (L. lactis) and human mesenchymal stem cells (hMSCs). L. lactis have been engineered to produce human C‐X‐C motif chemokine ligand 12, thrombopoietin, vascular cell adhesion protein 1, and the 7th–10th type III domains of human fibronectin, with the aim of recreating the native bone marrow conditions ex vivo. This active microenvironment has been shown to maintain key hMSC stemness markers, preventing their osteogenic and adipogenic differentiation, and maintaining high stem cell viability and physiological cell‐to‐substrate adhesion dynamics. This work presents proof of concept data that hMSC stemness can be regulated by living materials, using a system based on the symbiotic interaction between different engineered bacteria and mammalian cells.

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“…This cell line was engineered to express Sox-1/e GFP, which is a marker for neural precursor cells and facilitates its differentiation into a neural lineage in an in vitro system. In this present study, we mimic oxidative stress in the brain using glutamate excitotoxicity in neural cells derived from the 46C cell line using 4−/4+ protocol as previously described; this protocol successfully generated neural cells in vitro to produce a mixed culture of neuronal cells and glial cells [ 6 – 8 ]. It is predicted that both forms of vitamin E (TRF and α -TCP) would exert a neuroprotective effect against oxidative stress in neural cultures.…”

Section: Introductionmentioning

confidence: 99%

Vitamin E‐Mediated Modulation of Glutamate Receptor Expression in an Oxidative Stress Model of Neural Cells Derived from Embryonic Stem Cell Cultures

Jalil

Khaza’ai

Nordin

et al. 2017

Evidence-Based Complementary and Alternative Medicine

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Glutamate is the primary excitatory neurotransmitter in the central nervous system. Excessive concentrations of glutamate in the brain can be excitotoxic and cause oxidative stress, which is associated with Alzheimer's disease. In the present study, the effects of vitamin E in the form of tocotrienol-rich fraction (TRF) and alpha-tocopherol (α-TCP) in modulating the glutamate receptor and neuron injury markers in an in vitro model of oxidative stress in neural-derived embryonic stem (ES) cell cultures were elucidated. A transgenic mouse ES cell line (46C) was differentiated into a neural lineage in vitro via induction with retinoic acid. These cells were then subjected to oxidative stress with a significantly high concentration of glutamate. Measurement of reactive oxygen species (ROS) was performed after inducing glutamate excitotoxicity, and recovery from this toxicity in response to vitamin E was determined. The gene expression levels of glutamate receptors and neuron-specific enolase were elucidated using real-time PCR. The results reveal that neural cells derived from 46C cells and subjected to oxidative stress exhibit downregulation of NMDA, kainate receptor, and NSE after posttreatment with different concentrations of TRF and α-TCP, a sign of neurorecovery. Treatment of either TRF or α-TCP reduced the levels of ROS in neural cells subjected to glutamate-induced oxidative stress; these results indicated that vitamin E is a potent antioxidant.

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Formation of embryoid bodies from mouse embryonic stem cells cultured on silicon-coated surfaces

Cited by 6 publications

References 30 publications

An innovative flow cytometric approach for small-size platelet microparticles: Influence of calcium

An innovative flow cytometric approach for small-size platelet microparticles: Influence of calcium

Living Biointerfaces for the Maintenance of Mesenchymal Stem Cell Phenotypes

Vitamin E‐Mediated Modulation of Glutamate Receptor Expression in an Oxidative Stress Model of Neural Cells Derived from Embryonic Stem Cell Cultures

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