Figure 1

Cancer stem cells (CSCs) isolation and characterization from human ovarian SKOV3 cell line. (a) SKOV3 cells were labeled with FITC-conjugated anti-CD133 and PE-conjugated anti-CD44 antibodies, and analyzed by a flow cytometer as described in Materials and methods. (b) Immunofluorescence staining of CD133 and CD44 in CSCs and SKOV3 cell line. Cells were directly labeled using FITC-conjugated anti-CD133 (green) and PE-conjugated anti-CD44 (red) antibodies; nuclei were stained blue with 4′,6-diamidino-2-phenylindole (DAPI). (c) SOX2 protein expression was stained using FITC-conjugated anti-SOX2 and analyzed by Flow cytometry in SKOV3 cells, immediately after isolation and after 48 h incubation in medium supplemented by 2% fetal bovine serum (FBS). (d) SOX2 mRNA expression in CSCs was carried out by real-time RT-PCR and compared with that expression in SKOV3 cells. Data are expressed as mean ± SD of three independent experiments. Statistically significant differences are indicated as *p < 0.05. (e) Cells (3 × 104) were cultured in serum free media supplemented with 10 µM epidermal growth factor (EGF) and 10 µM basic fibroblast growth factor (bFGF) in non-treated six-well plates for seven days and spheroid formation was visualized by light microscopy in both CSCs and SKOV3 cells.