Isolation, growth, and characteristics of human ovarian surface epithelium

To isolate human ovarian surface epithelium (OSE) and studied its characteristicsin vitro, we used the scraping method with an enzymatic procedure. After incubation of the ovary in collagenase type 1 solution, the surface cells were removed by gentle scraping with surgical blade. Cells obtained as a cluster after unit gravity sedimentation with 5% bovine serum albumin in medium 199 were cultured in medium 199 containing 15% fetal bovine serum. The viable cell number in a single ovary was 0.1 to 2.7×10⁶, and the cell population doubling time was between 7 to 10 days. Confluent monolayers were formed after 13–20 days and subcultured one to three times. The monolayers mostly had a cobblestone appearance, and fusiform or polygonal cells were also observed. By cytochemistry, immunocytochemistry and scanning and transmission electron microscopy, the cells in short-term culture were shown to have characteristics of mesothelial OSE cells.