Effect of coconut water and Braun-Collins solutions at different temperatures and incubation times on the morphology of goat preantral follicles preserved in vitro

Abstract

Preservation of preantral follicles becomes very important to ensure follicle quality at the onset of cryopreservation or in vitro culture. However, for domestic animals, the ovarian donor of preantral follicles for in vitro studies is commonly encountered far away from reproduction laboratories. We investigated the effectiveness of coconut water and BraunCollins solutions on the preservation of goat preantral follicles. At the slaughterhouse, the ovarian pair of each animal was divided into 19 fragments. One ovarian fragment was immediately fixed (Control -Time 0). The other 18 fragments were randomly distributed into tubes containing 2 mL of coconut water or Braun-Collins solution at 4 °, 20 ° or 39 °C and then stored for 4, 12 or 24 h. Histological analysis showed that the storage of ovarian fragments in coconut water and Braun-Collins solutions at 20 ° or 39 °C for 12 or 24 h significantly reduced (P < 0.05) the percentage of morphologically normal preantral follicles when compared with the control. However, storage in coconut water at 20 °C for 4 h and in both solutions at 4 °C kept the percentage at control values. Ultrastructural analysis of follicles exposed to the stated conditions confirmed the integrity of preantral follicles stored at 4 °C in Braun-Collins and coconut water solutions for up to 12 and 24 h, respectively. Reduced cellular metabolism at 4 °C may explain why the best preservation of preantral follicles was at 4 °C, which may suggest a useful method for ovary transport in the future.

Effects of lowered temperatures and media on short-term preservation of zebu (Bos indicus) preantral ovarian follicles

“…Some experiments have been conducted to evaluate the effects of storage temperatures and media on the quality of preantral follicles [1] and [2]. In general, researchers found storage temperatures of 4 and 20 °C to be more suitable than 39 °C (body temperature) for the preservation of the normal morphology of sheep [3] and goat [1], [2] and [4] preantral follicles in situ.…”

“…In general, researchers found storage temperatures of 4 and 20 °C to be more suitable than 39 °C (body temperature) for the preservation of the normal morphology of sheep [3] and goat [1], [2] and [4] preantral follicles in situ. Storage at 20 °C has proved suitable for the preservation of preantral follicles for up to 4 h [1] and [4] and 4 °C was effective for 12 or 24 h of preservation [1], [2] and [4], depending on the storage media.…”

“…In general, researchers found storage temperatures of 4 and 20 °C to be more suitable than 39 °C (body temperature) for the preservation of the normal morphology of sheep [3] and goat [1], [2] and [4] preantral follicles in situ. Storage at 20 °C has proved suitable for the preservation of preantral follicles for up to 4 h [1] and [4] and 4 °C was effective for 12 or 24 h of preservation [1], [2] and [4], depending on the storage media. Media tested for the preservation of preantral follicles in ovarian pieces include saline solution (SS) [2] and [3], Braun-Collins solution [1], coconut water (CW) [1], [2] and [3], and M199 (with or without indole-3-acetic acid) [4].…”

“…Storage at 20 °C has proved suitable for the preservation of preantral follicles for up to 4 h [1] and [4] and 4 °C was effective for 12 or 24 h of preservation [1], [2] and [4], depending on the storage media. Media tested for the preservation of preantral follicles in ovarian pieces include saline solution (SS) [2] and [3], Braun-Collins solution [1], coconut water (CW) [1], [2] and [3], and M199 (with or without indole-3-acetic acid) [4]. However, all these studies were performed in small ruminant (sheep and goat) ovaries, and nothing is known about preserving preantral follicles in zebu cow ovaries.…”

“…The chosen temperatures (4 and 20 °C) were those that yielded the best results in previous studies [1], [2], [3] and [4], and the three storage times (6, 12, and 18 h) were taken at regular intervals of 6 h.…”