Figure 3

Immunohistochemical staining of 2D and 3D cultured FTSECs and primary fallopian tube tissue. Immunohistochemical staining of p53 (clone Do7) and a proliferation marker, MIB1, shows high expression of both markers in 2D cultured cells but low expression in 3D FTSEC cultures (at day 15), and fallopian tube epithelia in vivo. Laminin and vimentin are expressed in 2D cultured cells, 3D cultured cells and in vivo. Expression of fibronectin and collagen I was lower in 2D cultured cells than 3D cultures and fallopian tube epithelia in fresh tissue specimens. Collagen IV is predominantly expressed in the basal lamina (arrow) and stroma of fallopian tubes but was expressed at high levels by 2D cultured cells. Expression of this marker was low/absent in 3D cultured cells. * Fibronectin, collagen I and collagen IV were also examined in FTSEC01, expression patterns were highly similar to data for FTSEC283 and FTSEC03. Expression of collagen IV was restricted to the basal lamina and stroma. None of the markers examined were differentially expressed by ciliated and secretory fallopian tube epithelial cells in vivo. As our cell isolates were predominantly PAX8 positive (Figure 1), we do not expect ciliated cells to be present in the FTSEC spheroid cultures. Brown stain denotes positive antigen detection, cells are counterstained with eosin (blue). Light microscopy. Scale bars = 100 μm.