20 research outputs found
Side population analysis.
<p>(A) Cytometric analyses of the side population. The CD133<sup>+</sup> fraction includes a small subset (0.97%), expressing the characteristic profile of a side population at FACS. (B) ABCG2 expression in SAOS2 cell line, showing an evident positivity; the grey line indicates the isotype control.</p
Colony-forming efficiency of CD133<sup>+</sup> cells versus CD133− population<sup>a</sup> in SAOS2, MG63 and U2OS cell lines.
a<p>Results are the mean±standard devition of three experiments from different cases.</p>b<p>Represents the number of colonies with respect to the number of wells plated in experiments.</p>c<p>Ratio between the percentage of colonies formed by CD133+ versus CD133− cells.</p
CD133 expression in adherent cells and floating spheres.
<p>(A) Immunohistochemical analyses on adherent cells and (B) floating spheres showing the presence of CD133 antigen (<u>arrows</u>). (Original Magnification. ×100); (C) Immunofluorescence analysis on SAOS-2 for CD133 PE, cytoskeleton is stained with phalloidin-FITC, nucleus with DAPI (Original Magnification. ×400); (D) Immunoflurescence analysis on SAOS-2 spheres for CD133 PE after 24 hours in adhesion. (Original Magnification ×200); (E) Confocal analyses on adherent cells and (F) floating spheres confirming the presence of the CD133 antigen. (Original Magnification. ×400).</p
Spheres assay.
<p>(A) Sphere clusters formed by CD133<sup>+</sup> cells in semisolid medium after 24 hours (Original Magnification ×100); (B) CD133<sup>−</sup> cells in semisolid medium after 7 days, do not form spheres. (Original Magnification ×100); (C) Sphere clusters formed by CD133<sup>+</sup> cells after 48 hours (Original Magnification ×200); (D) Sphere clusters formed by CD133<sup>+</sup> cells after a new sorting (Original Magnification ×400).</p
Cytometric analyses for CD133 on SAOS2, MG63 and U2OS cell lines.
<p>A CD133<sup>+</sup> cell population can be detected in all the three cell lines.</p
Cytometric analyses for CD133 and OCT3/4 on sarcospheres in SAOS2, MG63 and U2OS.
<p>The blue line indicates isotype controls, red and green lines indicate the expression of CD133 at the 4<sup>th</sup> and 6<sup>th</sup> cell passage, respectively. In the histograms, OCT3/4 expression is analyzed at the 6<sup>th</sup> cell passage; the green line indicates isotype controls. Sarcospheres both in CD133 and OCT3/4 are strongly positive.</p
Cytometric analyses for CD133, CD44, CD90, CD34 and CD29 on SAOS2, MG63 and U2OS cell lines.
<p>All three cell lines are negative for CD34 antigen but they evidence equal strong positivity for CD29, CD44 and CD90.</p
CD133 intracellular expression in adherent cells.
<p>(A) Figure performed at FACS showing intracellular expression of CD133 antigen in SAOS-2, MG-63 and U2OS. (B) Figure showing at the Real time PCR the mRNA transcript expression in CD133<sup>+</sup> and CD133<sup>−</sup> cells. The levels are almost identical as detected by in both CD133 positive and negative cells.</p
Cell cycle, proliferation and growth analyses.
<p>(A) Figure of cell cycle analyses performed on SAOS2, MG63 and U2OS cell lines. A CD133<sup>+</sup> cell population is mainly observable in the G2\M phase, whereas CD133<sup>−</sup> cells were predominantly in G0\G1; (B) Figure showing Ki-67 reactivity. CD133<sup>+</sup> cells resulted to be Ki-67<sup>+</sup>, whereas CD133<sup>−</sup> were mainly negative for this marker; (C) Figure showing growth curves of CD133<sup>+</sup> cells with respect CD133<sup>−</sup> cells in SAOS2, MG63 and U2OS cell lines. CD133<sup>+</sup> cells possess a high proliferative potential in all the three cell lines.</p
Spheres formation, cytometric analysis and telomerase activity.
<p>(A) Sphere clusters formed by CD34<sup>+</sup>/CD90<sup>+</sup> cells in semisolid medium after 24 hours (Original Magnification×100); (B) Cytometric analysis on adherent cells for CD90 (80%) and CD34 (80%) antigens and on floating spheres for CD90 (2–3%) and CD34 (95–98%) antigens; (C) Telomerase activity of differentiated endothelial cells (ΔA = 0.160) was significantly reduced (p<0.001) respect to undifferentiated CD34<sup>+</sup>/CD90<sup>+</sup> cells (ΔA = 0.377)</p