Morphological appearance of MCTS culture of MCF-7.

<p>(A) With the facilitation by centrifugal force, MCF-7 cells were organized into a three-dimensional multicellular spheroidal structure. Under phase contrast microscope, the structure appeared to be compact and the cells were rigidly integrated into the solid structure where individual cells were indistinguishable from each other (magnification: 40x, scale bar: 100 µm). (B) Appearance of the culture under scanning electron microscopy (magnification: 160x, scale bar: 100 µm). (C) Single cells within the spheroid culture were connected to adjacent cells through cell-cell junction (arrow), which were responsible for the densely packed organization of the cells (magnification: 4000x, scale bar: 5 µm).</p

Viability of the monolayer and MCTS cultures of MCF-7 determined from the MTT assay.

<p>Monolayer and MCTS cultures of the MCF-7 cell line were exposed to tamoxifen for 4 days. MTT assay for the MCTS cultures was carried out with some modification to the standard protocol by Mosmann (1983) <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0044640#pone.0044640-Mosmann1" target="_blank">[11]</a> while assay for the monolayer culture was performed in accordance to the original method.</p

Induction of apoptosis in MCTS cultures of MCF-7 after 4-day exposure to different concentrations of tamoxifen.

*<p>Statistical significance (P<0.05) between control cells and treatment groups.</p

Comparison of the cytotoxicity increment in MCTS cultures assessed by MTT and LDH release assays.

<p>Relative to the untreated control, which was normalized to 0% in both assays, reduction in cell viability of treated samples assessed by MTT assay was compared to the elevation in cytotoxicity determined from the LDH release assay. The results were presented as means ± S.E.M. of three independent experiments.</p

Cytotoxicity of tamoxifen citrate against monolayer and MCTS cultures of MCF-7.

<p>Cytotoxicity was determined by quantifying the percentage of LDH release relative to the maximum LDH release of untreated spheroid cultures. The results were presented as means ± S.E.M. of three independent experiments.</p

Muticellular tumor spheroidal culture of MCF-7 with homogenous sizes.

<p>The cells were cultured in a 96-well plate that was coated with agar gelrite. Cell aggregation was facilitated by centrifugation without the addition of any extracellular matrix to induce the formation of spheroid (Magnification: 1x, scale bar: 1 cm).</p

Proliferation index of emigrant cells compared to pre-cultivation mixtures of proventriculus, splenocytes and intestine or dissociated agglomerates obtained using a BrdU ELISA kit.

<p>The values were the means ± SEM of three experiments.</p

Double staining immunophenotyping of IgM⁺ and Bu-1a-F⁺ subpopulations in preculture mixtures, agglomerates and emigrant cells.

<p>The values were the mean percentages of total cell ± SEM of three experiments.</p

The age of chick embryos (days) and the number of successful replicates per number of attempts.

<p>Each replicate used one embryonic chicken and 6 culture attempts to form agglomerates were set up in each case. The formation of at least 5 out of 6 agglomerates was required for the scoring as successful. When 19 and 20 day chicks were used, agglomerates were not formed. Each replicate used one embryonic chicken, and six mini organs were attempted for each replicate. At least 5 out of six mini organs must be formed for the attempt to count as a successful replicate. For day 19 and 20, none of the mini organs was successfully formed in all three replicates.</p

Electron micrograph of agglomerate infected with NDV virus strain AF2240.

<p>(A) Agglomerate infected for 24 hours. Arrows indicate spherical virus particles in vesicles and cytoplasm of the cells. (B) 5000× magnification of NDV particles. Note spherical particles 20–28 nm in diameter detected in the agglomerate. (C) Note virus particles in the mitochondria and damage to their structure.</p