Figure 3

Cell growth rate and tumorigenicity assays. A) Growth curves of the three new cell lines as well as a previously characterized EOC TOV-112D cell line. 100 000 cells were plated onto 60 mm petri dishes. Cells were trypsinised and counted every 48 h for two weeks. Experiments were performed two times in triplicate. B) Wound-healing assay of the same four cell lines. Cells were plated onto a 12 well dish and at near confluence wounds were generated. Cells were methanol fixed and treated with Giemsa Stain at different time points in order to evaluate cell migration (0 h, 8 h, 24 h and 48 h after the scratch was performed). The experiments were performed twice in triplicate. C) Invasion assay using modified Boyden chambers. The capability of the cells to invade through matrigel membranes was verified and the invasion potential increased from TOV-112D, OV-1946, TOV-2223 to TOV-1946. The experiments were performed in duplicate. D) The capacity of the cells to form three-dimensional structures in hanging droplets was monitored. TOV-112D cells were able to form very compact spheroids, OV-1946 formed less compact spheroids, TOV-1946 cells formed loose aggregates of cells and TOV-2223 cells were unable to form any 3-D structure. Spheroid formation capability was visualized after four days.