Culture and characterization of normal ovarian epithelial and fallopian tube epithelial cells.

<p><b>A</b>, Replicate plates of normal fallopian tube epithelial cells were cultured in WIT-fo medium (green and purple lines) or standard medium (red, grey lines, 1:1 mixture of Dulbecco’s Modified Eagle’s Medium (DMEM) and Ham’s F12, supplemented with 0.1% BSA, 5% serum). In WIT-fo medium, normal fallopian tube epithelial cells from two different patients divided continuously for at least 30 days and > 100 days and reached at least 12 and >20 population doublings respectively (green and purple lines). In contrast, matched cells from the same donors growth arrested in the DMEM/Ham’s F12 medium (red, grey lines). Normal fallopian tube epithelial cells were isolated from additional patients 3 and 4. <b>B</b>, Normal ovarian epithelial cells were cultured in WIT-fo medium (purple and blue lines) or standard medium (red line, MCDB 105/Medium 199 (M199) (1∶1 mixture) with 10% FBS and 2 mm l-glutamine). Primary normal ovarian epithelial cells were from patients 2 and 3 (cells cultured in standard medium were from patient 3). Matched cells from patient 3 growth arrested in the MCDB105/M199 medium (red line). <b>C-D</b>, Normal human ovarian tissue; immunoperoxidase staining of formalin-fixed paraffin embedded (FFPE) sections with PAX8 demonstrates that ovarian inclusion cyst epithelium is PAX8⁺ (brown nuclear stain) (<b>C-D</b>) while ovarian surface epithelium is in general PAX8 negative (<b>C</b>), rare presence of rare Pax8 positive cells have been reposted on the ovarian surface. <b>E,</b> Normal human fallopian tube tissue; double immunoperoxidase staining of FFPE sections shows ciliated cells are FOXJ1⁺ (nuclear brown) and non-ciliated cells are PAX8⁺ (nuclear red). <b>F,</b> Normal human fallopian tube tissue; double immunoperoxidase staining of FFPE sections shows that ciliated cells are FOXJ1⁺ (nuclear brown) and non-ciliated cells are CK7⁺ (cytoplasmic red) (scale bar (C-F) = 20 µM). <b>G,</b> Summary of cell type specific characterization markers.