In vitro fertilization of horse follicular oocytes matured in vitro

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Abstract
In vitro fertilizing ability of stallion spermatozoa was assessed using horse follicular oocytes matured in vitro. After collection, stallion spermatozoa were either: 1) washed and incubated in TALP medium with 3 mg/ml bovine serum albumin (BSA) and 10 μg/ml heparin for 4 h, 2) washed and incubated in TALP with 3 mg/ml BSA for 3 h and cultured for a further 1 h with 1 mM caffeine and 5 mM dbcAMP, 3) washed and incubated in TALP medium with 3 mg/ml BSA at pH 7.9–8.2 for 2–4 h, or 4) diluted and incubated in TALP medium with 10 mg/ml BSA and 7.14 μM calcium ionophore A 23187 for 5–10 min followed by washing. After a given pretreatment, suspensions were diluted into B2 medium to a concentration of 5 × 106 sperm/ml and co‐incubated with oocytes for 12 h or 24–48 h. In the ionophoretreated group, 18 of 54 oocytes (33%) were fertilized by 12 h, and 11 of 45 (24%) cleaved by 24–48 h. Evidence of fertilization was not found in the oocytes incubated with spermatozoa from other treatment procedures.