GROWTH OF NORMAL HUMAN OVARIAN SURFACE EPITHELIAL-CELLS IN REDUCED-SERUM AND SERUM-FREE MEDIA

The human ovarian surface epithelium (OSE) is believed responsible for over 85% of ovarian cancers, yet little is known about the normal bio logy of these cells. To date, culture of OSE has only been reported in media with high serum supplements. We have developed two media, one w ith less than 1% of serum (OSEM-1) and the other comprised of highly p urified and defined materials (OSEM-2), which allow us to study OSE un der relatively defined conditions. By substituting 0.05% of Pedersen's fetuin for 15% fetal bovine serum (FBS) with Medium 199/MCDB105 basal medium, the cell numbers reached 50 to 60% of those in the presence o f 15% FBS over 7 days. However, over several weeks, the total number o f population doublings achieved were comparable to those in 15% FBS. A ddition of insulin, transferrin, ethanolamine, lipoic acid, and phosph atidylcholine to the medium with Pedersen's fetuin (OSEM-1) enhanced g rowth up to 20% more than in their absence. Supplementation of M199/10 5 with highly purified (>99%) fetuin, alpha2-macroglobulin, and hydroc ortisone resulted in a defined medium (OSEM-2) that permitted 1 to 2 d oublings/7 days. In addition, cells maintained a more normal, epitheli al-like morphology in culture for a longer period in the presence of P edersen's or purified fetuin than in M199/105/15% FBS, thus increasing the number of morphologically normal cells available for experimentat ion. Addition of 0.05% Pedersen's fetuin to M199/105 in the presence o f 6 to 8% FBS resulted in levels of growth equivalent to those in M199 /105/15% FBS alone. We are now able to study the effects of various co mpounds on the growth and differentiation of OSE under defined conditi ons, and have reduced the requirement for FBS to produce large numbers of OSE cells.