Distinct incubation for homologous in vitro spermatozoa binding on swine oocytes subjected to different storage conditions

The sperm in vitro binding assay in homologous oocytes can be used to estimate the boar fertility potential, but its usefulness may be limited by laboratorial structure and oocytes availability. This study aimed at determining the effect of distinct methods of oocytes conditioning and incubation media for the in vitro penetration (IVP) test. Oocytes used in the IVP test were: fresh and conditioned in PBS (T1); cooled and conditioned in PBS at 5 °C for 48 h (T2); or stored in ovaries frozen at −20 °C (T3). For each treatment, two incubation media were tested at 39 °C for 6 h: modified TRIS buffer medium (mTBM); or Beltsville Thawing Solution (BTS) extender. The responses of interest were: IVP and polyspermy rates; and the number of penetrating spermatozoa per oocyte. All responses observed with incubation in BTS were inferior to those observed with incubation in mTBM (P<0.0001). When incubation was done in mTBM, none of the responses differed across treatments (P>0.05). However, when incubation was in BTS, all the three responses were superior for T1 than for T2 and T3 (P<0.05). Thus, the IVP test may be conducted with ovaries either cooled or recovered from frozen ovaries with results similar to those observed with fresh oocytes, if incubation is done in mTBM.