Survival and growth of XXXX primordial follicles after in vitro culture of ovarian cortical slices in media containing coconut water.

@entity11896 @entity1160 @entity3625 @entity4259 @entity72368 @entity72369 @entity528 @entity1237 @entity2388
The development of culture systems to support the initiation of growth of primordial follicles is important to the study of the factors that control the earliest stages of folliculogenesis. We investigated the effectiveness of five culture media, two supplements and three culture periods on the survival and growth of @entity1237 primordial follicles after culturing ovarian cortex. The media were based on minimal essential minimum (MEM) and @entity11896 ( @entity11896 ) added in the proportion of 0, 25, 50, 75 or 100%. The two supplements were none versus supplemented with insulin- @entity72368 - @entity4259 , @entity2388 , @entity1160 , @entity3625 , and BSA. Pieces of @entity1237 ovarian cortex were cultured in the media for 1, 3 or 5 days and representative samples were evaluated at day 0 as non-cultured controls. The replicates were the two ovaries of five mixed breed @entity1237 . The number of primordial, intermediate, primary and secondary follicles at each period of culture and the number of degenerated follicles were evaluated. Mitotic activity of granulosa cells was studied by immunolocalization of @entity72369 ( @entity72369 ). The number of follicles in each stage and degenerated follicles were statistically analyzed by ANOVA using a factorial design and the significance of differences assessed using Tukey test. Chi-square test was used to compare the percentage of follicles with @entity72369 positive granulosa cells. As the culture period progressed, the number of primordial follicles fell and there was a significant increase in the number of primary follicles. The fall in the number of primordial follicles was particularly marked after 1 day culture. No effect of media on the number of primordial and primary follicles was observed after culture, but MEM as well as supplements increased the number of intermediate follicles. @entity528 was kept at the same level after culture in the media tested, except for pure @entity11896 that increased the number of degenerated follicles. In contrast, addition of supplements to culture media reduced follicular degeneration. In non-cultured tissue, @entity72369 was expressed in granulosa cells of 31.6% of the growing follicles. This percentage had not significantly changed after 5 days culture in the various media, indicating the maintenance of proliferation activity of granulosa cells during culture. In conclusion, it is shown that @entity1237 primordial follicles may be successfully activated after in vitro culture in all media tested. However, when pure @entity11896 is used the follicular degeneration is enhanced, but the addition of supplements to culture media decrease follicular degeneration.
