Normal Development Following In Vitro Fertilization in the Cow

Abstract

A repeatable procedure for fertilization of bovine ova in vitro is described. Oocytes were recovered from ovarian follicles or from oviducts near the time of ovulation following treatment of donors with pregnant mare's serum gonadotropin (PMSG) and prostaglandin F2 alpha (PGF2 alpha). For in vitro capacitation semen was incubated, then high ionic strength treated and subsequently incubated in defined medium prior to insemination of oocytes. In one experiment frozen bull semen was successfully used. In experiments with 4 bulls (B, C, D, F), 34 (43.6%) of 78 ova and 13 (19.7%) of 66 follicular oocytes were fertilized in vitro. In the last series (spermatozoa from Bull F) the fertilization of 22 (62.9%) of 35 tubal ova was achieved. In vitro development proceeded to the 8-cell stage. No fertilization in vitro followed use of one male (Bull E), even though his spermatozoa could penetrate zona-free hamster ova in vitro, and higher than usual bacterial contamination of his semen was implicated as the probable cause. Findings suggested vigorous progressive sperm motility and acrosome integrity to be important features of good sperm samples. In one experiment a 4-cell stage embryo was transferred with the result that the recipient gave birth to a normal bull calf on June 9, 1981. The first calf resulting from in vitro fertilization has been found to be completely normal.

Pregnancies after co-culture of cumulus cells with bovine embryos derived from in-vitro fertilization of in-vitro matured follicular oocytes

“…In-vitro fertilization of cow ova began in the 1970s (Edwards, 1973;Bregulla et ai, 1974;Iritani & Niwa, 1977;Brackett et al, 1978) and, in 1981, the first calf was born after in-vitro fertilization (Brackett et al, 1982). Although other calves have since been born (Brackett et al, 1984), the technology used has been considered unsuitable for field application.…”

“…In-vitro fertilization of oocytes would have a major impact on bovine embryo transfer (Brackett et al, 1982;Brackett, 1983). A wide variety of culture media has been used but only limited in-vitro development of preimplantation cow embryos has been reported (Brock & Rowson, 1952;Thibault, 1966;Brinster, 1968;Sreenan, 1968;Sreenan et al, 1968;Onuma & Foote, 1969;Seidel et ai, 1971;McKenzie & Kenney, 1973).…”

Capacitation in the presence of methyl-β-cyclodextrin results in enhanced zona pellucida-binding ability of stallion spermatozoa

“…In contrast to the considerable IVF successes that have been achieved in other mammalian species such as mice (Whittingham 1968, Mukherjee & Cohen 1970 and cattle (Iritani & Niwa 1977, Brackett et al 1982, the development of a robust IVF system for equine species remains difficult; only two foals having been reported using conventional IVF protocols (Palmer et al 1991, Bezard 1992). The fundamental impediments, which are at least partly responsible for the slow progression of equine IVF, include the limited availability of equine oocytes and the slow development of in vitro oocyte maturation.…”

Effect of heparin on the expression of calmodulin-binding proteins in bull spermatozoa

“…Successful in-vitro fertilization has been achieved in cattle following capacitation treatment of spermatozoa with a hyperosmotic medium (Brackett et al, 1982;Lambert et al, 1986) or with the glycosaminoglycan heparin (Parrish et al, 1985(Parrish et al, , 1986a(Parrish et al, ,b, 1988. The molecular mechanisms are still not elucidated, but Ca2 + is recognized as one of the most important factors involved in the process of capacitation (Fraser, 1982(Fraser, , 1987.…”