Figure 3.

OCSCs spheroids were obtained with ALDEFLUOR assay and FACS. (A) The microscope image of irregular spheroids derived with the A2780 cells with serum free culture for three weeks. (B) The spheroids were sorted with FACS analyses of ALDH activity with ALDEFLOUR staining in the presence or absence 5 μL of diethylaminobenzaldehyde (DEAB) reagent, which was the ALDH1 inhibitor. The ALDH1⁺ (54.8%) was sorted by flow cytometry compared with Control groups (1.07%) (p< .01). After sorting, the ALDH1⁺ OCSCs and ALDH1^– A2780 cells were cultured with serum-free culture. (C) One month later, ALDH1⁺ OCSC spheroids were full and globular. (D) The ALDH1⁺ OCSC spheroids changed into monolayers with serum culture medium for 48 h (scale bar = 100 μm). (E, F) Compared with ALDH1^– A2780 cells, the ALDH1⁺ OCSC spheroids had higher tumorigenesis abilities in vivo (p< .01).