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Experimental Results on Cellular and Subcellular Systems Exposed to Low-Frequency and Static Magnetic Fields
Published in Ben Greenebaum, Frank Barnes, Biological and Medical Aspects of Electromagnetic Fields, 2018
Myrtill Simkó, Mats-Olof Mattsson
Further studies of gene expression as a marker for differentiation have been performed. Thus, a global mRNA profiling study on human embryoid body derived lymphatic vascular endothelial cells (LVEC) (a stem cell line) showed that MF at 0.23–0.28 T for 24 h affected expression levels of hundreds of genes (Wang et al. 2009). The authors identified nine different signaling networks that were directly involved and were selected to further characterize effects linked to the inflammatory IL-6 pathway. Proliferation was suppressed by the exposure, but there were no indications of any toxic effects. Microscopic investigations showed that the cells developed dendrite-like outgrowths, which can be found in both neural and glial precursors. However, on the protein levels, the cells increased their expression levels of vimentin and Gal-C, suggesting that the exposure had stimulated the stem cells to develop along an oligodendrocyte pathway, whereas markers for neurons and for astrocytes were downregulated. That the IL-6 signaling network was induced suggests that the initial events are effects of changing Ca2+ levels, since IL-6 signaling is modulated by Ca2+ influx.
Proliferation of mouse embryonic stem cells on substrate coated with intact silkworm sericin
Published in The Journal of The Textile Institute, 2022
Chihiro Umehara, Ai Ai Lian, Yuichiro Funahashi, Keiko Takaki, Rina Maruta, Yuto Ohmaru, Yoko Okahisa, Takashi Aoki, Hajime Mori, Eiji Kotani
To induce the myocardial differentiation of EB5-αMHC-DsRed cells, the differentiation medium (MEM-α; Gibco) supplemented with 10% (v/v) FBS, 2mM L-glutamine (Wako Chemicals), 0.1mM NEAA, 1mM sodium, 0.1mM 2-ME, and 20ng/mL blasticidin was used. EB5-αMHC-DsRed cells (1.0×105 cells/well) were cultured on the gelatin- or sericin-coated substrate in the wells of a 6-well plate containing 2mL of differentiation medium supplemented with 10ng/mL rhLIF and 150ng/mL recombinant mouse Noggin (rmNoggin). After cultivation for 3days, the EB5-αMHC-DsRed cells were recovered and resuspended in 100µL of differentiation medium containing 150ng/mL rmNoggin and transferred to a flat-bottom 96-well plate (Nunc) (6,000 cells/well). The cells were further incubated for 5days for embryoid body formation.
Preparation of cell aggregates incorporating gelatin hydrogel microspheres containing bone morphogenic protein-2 with different degradabilities
Published in Journal of Biomaterials Science, Polymer Edition, 2018
Shuhei Tajima, Yasuhiko Tabata
Recently, three-dimensional (3D) cell culture technologies have been increasing noticed [1–4]. The conventional cell culture has been performed in two-dimensional (2D) systems, which is quite different from the local environment of cells in living tissues. Looking at the structure of body tissues, such as liver and bone, cell aggregates physiologically work as the minimum unit of function [5]. In the 3D cell culture, it is possible that the cell aggregation permits efficient cell–cell interactions, resulting in an enhanced biological functions of cells. For example, embryonic stem cells generally aggregate to form an embryoid body, and consequently initiate differentiation into different cell lineages [6]. The aggregation of liver cells to form a spheroid is necessary to enhance their metabolic activity [7]. In addition, cell aggregates produce extracellular matrix proteins more efficiently than single cells [8]. However, there are some problems for the culture of cell aggregates. One of them is that when the cell aggregate grows up, cells in the center of large aggregates die because of the lack of oxygen and nutrients supplies [9]. The previous study revealed that the incorporation of slow-degraded GM prevents the mouse preosteoblast MC3T3-E1 cells aggregated from a lack of oxygen in the long-term culture because oxygen and nutrients can be permeated through the water phase of hydrogel matrix [10], resulting in enhanced cell proliferation and osteogenic differentiation in MC3T3-E1 aggregates [11]. The number of papers about the 3D cell aggregates culture system combined with microspheres has been increased recently [12–16]. However, there are few research reports to promote and control the functions of cells in cell aggregates by making use of microspheres.