Production of mitogenic factors by cell types of bovine large estrogen-active and estrogen-inactive follicles

This study investigated whether large follicles (estrogen-active and estrogen-inactive) of cows produce factors with mitogenic activity. Large, preovulatory follicles (greater than or equal to 9 mm in diameter) were classified as estrogen-active or -inactive based on ratio of estrogen: progesterone concentrations in follicular fluid. After incubation of granulosa cells and thecal tissues from follicles, granulosa cell conditioned media (GCM), thecal conditioned media (TCM) and follicular fluid (FFL) were evaluated for effects on proliferation of bovine aortic endothelial (BAE) and BALB/3T3 (3T3) cells. Pools of GCM, TCM and FFL stimulated proliferation of BAE and 3T3 in a dose-dependent fashion. Across all follicles (n = 20), GCM had greater stimulatory effect on proliferation of BAE than on proliferation of 3T3 (135 vs 115% of unconditioned media controls), whereas TCM stimulated proliferation of BAE and 3T3 to a similar extent (128 and 128%). Across type (GCM and TCM) of conditioned media, estrogen-active follicles stimulated proliferation of BAE more than proliferation of 3T3 (137 vs 121% of unconditioned media controls), whereas estrogen-inactive follicles stimulated proliferation of BAE and 3T3 to a similar extent (120 vs 122%). As observed for GCM, FFL across all follicles had a greater stimulatory effect on proliferation of BAE than on proliferation of 3T3 (159 vs 141%). Granulosa-conditioned media stimulated proliferation of BAE and 3T3 only when obtained from estrogen-active follicles; mitogenic activities of TCM and FFL were not influenced by type of follicle. These data demonstrate that granulosa cells of large preovulatory bovine follicles secrete a mitogenic factor(s) that is more stimulatory for proliferation of BAE than for 3T3.(ABSTRACT TRUNCATED AT 250 WORDS)