Abstract
The role of follicle cells in the acrosome reaction of frozen‐thawed bovine spermatozoa, in vitro fertilization, cleavage, and development in vitro was investigated. Cumulus‐oocyte complexes were cocultured and matured in vitro with additional granulosa cells for 24 hr. Immediately before in vitro insemination, the oocytes were divided into three types with different follicle cells: denuded and corona‐ and cumulus‐enclosed oocytes. The proportion of live, acrosome‐reacted spermatozoa significantly increased at 3 and 6 hr after insemination in all types of oocytes. However, the mean proportion of live, acrosome‐reacted spermatozoa that inseminated cumulus‐enclosed oocytes at 6 hr after insemination was significantly higher than that of spermatozoa inseminating denuded oocytes (18.3% and 13.3% respectively). The frequency of in vitro fertilization was significantly higher for cumulus‐enclosed oocytes (65.4%) than for denuded and corona‐enclosed oocytes (30.8% and 39.4%, respectively). Cumulus‐enclosed oocytes when cocultured with oviduct epithelial cells also had significantly higher rates of cleavage (two‐ to eight‐cell, 59.8%; eight‐cell, 22.4%) and blastocyst formation (7.7%) than denuded and corona‐enclosed oocytes. No eight‐cell embryos or more advanced stages of embryonic development were observed in either denuded or corona‐enclosed oocytes without the coculture. The present results indicate that cumulus cells at fertilization play an important role in inducing the acrosome reaction and promoting a high fertilization rate, cleavage, and development into blastocysts in vitro.