Fig. 4
OC lineage-specific differentiation potency. A Schematic of the random differentiation procedure and determination of lineage-specific differentiation abilities in vitro. IF staining was performed with CK7 and CA125 antibodies under both nondifferentiation and differentiation conditions in iOVCAR-3-OSKM cells. Scale bars: 50 µm. B Determination of lineage-specific differentiation in vivo. IHC staining was used to assess tumors derived from iOVCAR-3-OSKM cells in NOD-SCID mice. Xenografts of iOVCAR-3-OSKM cells showed both CK7 and CA125 expression. The upper panels show IHC images at 40X magnification (scale bars: 250 µm), and the lower panels show IHC images at 200X magnification (scale bars: 50 µm). C Schematic of the organoid culture procedure. A total of 3 × 105 single cells were suspended in medium containing 2% Matrigel with 50 nM E2 and seeded onto culture dishes coated with 100% Matrigel for organoid 3D culture for two weeks or four weeks. The number (> 20 µm organoids were counted under 100X magnification) and diameter (10 organoids were measured under 200X magnification) of two-week organoids were calculated. Scale bars (from left to right): 250 µm; 100 µm; 50 µm. Error bars indicate SD. *P < 0.05, Student’s t test. D Two-week organoids were fixed, and IF staining was performed. iOVCAR-3-OSKM cells formed organoids and expressed CK-7 and CA125. Scale bars: 50 µm. E H&E staining of xenografts and four-week organoid cultures. Xenograft tumors and organoids were collected and embedded in paraffin. Sections were stained with H&E. Scale bars: 25 µm