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Fig. 1 | Journal of Biomedical Science

Fig. 1

From: Suberoylanilide hydroxamic acid represses glioma stem-like cells

Fig. 1

Serum-free suspension culture induces the enrichment of Glioma stem-like cells (GSCs). a and b GSCs from U87MG GBM cells were grown in serum-free DMEM/F12 medium/suspension culture for 2–14 days. Sphere morphology (2 weeks) was photographed by light microscopy and spheroids (2–14 days) were harvested and subjected to Western blotting using indicated antibodies. c The protein expression levels were quantified by imageLab software 5.2.1 (Bio-RAD). The protein expression of U87MG parental (Adherent) was defined as 1 and other values were accordingly normalized. d and e U87MG spheres were reattached to without poly-HEMA-coated plates in a serum-containing medium for 1 day, in order to induce GSCs differentiation. The levels of stem cell-related markers and Vimentin, an astrocytic marker, in parental cells (Adh), GSCs (Sph), and differentiated GSCs (Dif) were measured via Western blot with antibodies as indicated (d). Representative images are shown in (e). f U87MG adherent cells and dissociated spheroids by trypsin/EDTA were plated at a density of 3 x 102 cells/well. After 0, 3, 6, and 9 days of culture, cell proliferation capacity was determined by using MTT assay. Bar graphs represent means ± standard errors of the means (SEM) from three independent experiments (t-test: *p < 0.05; **p < 0.01). g A single cell isolated from primary spheroids was cultured in a 96-well dish. The propagation of a secondary spheroid formation was recorded at day 0, 3, and 6, separately (upper panels). Subsequently, this secondary spheroid was reattached to plates to induce cellular differentiation (lower panel). h Immunofluorescence images of CD133 staining in the secondary spheroid and differentiated cells. The nuclei were stained with DAPI (blue)

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