Effects of two treatments on semen from different bulls on in vitro fertilization results of bovine oocytes

Aoyagi, Yoshito; Fujii, K.; Iwazumi, Yasuaki; Furudate, Makoto; Fukui, Yutaka; Ono, Hitoshi

doi:10.1016/s0093-691x(88)80060-8

Cited by 32 publications

(14 citation statements)

References 21 publications

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“…In bovine IVF systems, both caffeine and theophylline have been used to induce sperm capacitation [11,[20][21][22][23]. Although theophylline has a greater ability to inhibit PDE than does caffeine [15], only caffeine has been applied for a majority of current porcine IVF systems [2, 3, 6-8, 12, 24, 25].…”

Section: Discussionmentioning

confidence: 99%

Production of Piglets Derived from In Vitro-Produced Blastocysts Fertilized and Cultured in Chemically Defined Media: Effects of Theophylline, Adenosine, and Cysteine During In Vitro Fertilization1

Yoshioka

Suzuki

Itoh

et al. 2003

Biology of Reproduction

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To further develop defined conditions for in vitro fertilization (IVF) and in vitro culture (IVC) of in vitro-matured porcine oocytes, we evaluated the effects of theophylline, adenosine, and cysteine in a chemically defined medium during IVF. Viability to full term of in vitro-produced blastocysts after IVF and IVC in chemically defined medium was also investigated by embryo transfer to recipients. A chemically defined medium, porcine gamate medium (PGM), was modified from porcine zygote medium (PZM-4), which was previously established. PGM was used as a basal medium for IVF and PZM-4 was for the culture of presumptive zygotes. Addition of 2.5 mM theophylline to PGM significantly increased the percentage of male pronuclear formation compared with controls (no addition). Addition of 1 microM adenosine to PGM supplemented either with or without 2.5 mM theophylline significantly reduced the number of penetrated spermatozoa compared with controls (no addition of adenosine). Supplementation with 0.2 microM cysteine in PGM containing both 2.5 mM theophylline and 1 microM adenosine further increased the percentage of development to the blastocyst stage, compared with no supplementation of cysteine, but there was no difference in fertilization parameters, such as monospermy and pronuclear formation, regardless of presence or absence of theophylline and adenosine. When Day 5 blastocysts were transferred into four recipients (20-25 blastocysts per recipient), all recipients became pregnant and farrowed a total of 21 live piglets. The present results clearly demonstrate that porcine blastocysts can be produced by IVF and IVC in chemically defined media and that they can develop to full term after embryo transfer.

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Section: Discussionmentioning

confidence: 99%

Production of Piglets Derived from In Vitro-Produced Blastocysts Fertilized and Cultured in Chemically Defined Media: Effects of Theophylline, Adenosine, and Cysteine During In Vitro Fertilization1

Yoshioka

Suzuki

Itoh

et al. 2003

Biology of Reproduction

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“…Various chemicals such as caffeine [1,2], heparin [3,4], calcium ionophore A23187 [5], and caffeine plus heparin [6] or caffeine plus calcium ionophore A23187 [7] have been used to induce capacitation of bovine spermatozoa in vitro. However, BSA is always included in media used for in vitro fertilization in these experiments.…”

Section: Introductionmentioning

confidence: 99%

In Vitro Fertilization of Bovine Oocytes in a Chemically Defined, Protein-Free Medium Varying the Bicarbonate Concentration1

Tajik

Wang

Okuda

et al. 1994

Biology of Reproduction

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Bovine cumulus-enclosed oocytes were matured in culture, freed from cumulus cells, and inseminated with frozen-thawed spermatozoa in a chemically defined protein-free medium containing 5 mM caffeine and 10 micrograms/ml heparin. No penetration of oocytes was observed in the medium without polyvinyl-alcohol (PVA); but when the medium was supplemented with 0.1-5 mg/ml PVA, penetration rates (9-16%) significantly increased. Sperm motility was also stimulated during incubation for 2 h in the presence of PVA. In the medium with 1 mg/ml PVA, a high penetration rate (24 of 62 = 39%) was observed at a sperm concentration of 10 x 10(6) cells/ml. When the bicarbonate concentration was changed in the fertilization medium containing 1 mg/ml PVA and 10 x 10(6) spermatozoa/ml, a high penetration rate (47 of 67 = 70%) and a high proportion (44 of 47 = 94%) of oocytes in which male and female pronuclei had developed were obtained at 46 mM NaHCO3. However, the penetration rate (58-95%), the incidence of pronuclear formation (64-96%), and the proportion of polyspermy (9-21%) varied according to the animal (five different bulls). Spermatozoa obtained from two bulls started to penetrate oocytes 5 h after insemination in the presence of 46 mM NaHCO3. This is the first report indicating that induction of capacitation of bull spermatozoa and penetration of oocytes matured in culture are possible in a chemically defined, protein-free medium.

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“…However, the applied modification limiting preparation of these fractions only to a centrifugation of thawed sperm did not allow removing dead spermatozoa that had been damaged following the cryoconservation procedure, which might exert a detrimental effect on fertilization results. Several authors dealing with in vitro embryo production in cattle (MILLER and HUNTER, 1987;AOYAGI et al, 1988;EYESTONE and FIRST, 1989;GALLI and LAZZARI, 1996;KĄTSKA et al, 1996;CHAMBERLAND et al, 2001;GALLI et al, 2003;KĄTSKA-KSIĄŻKIEWICZ et al, 2005) have described the variability among bulls in the ability of their sperm to become capacitated, fertilize oocytes and subsequently produce competent embryos developing to the blastocyst stage. In terms of factors affecting the outcome of IVF, bull variability has been the main source of difficulty in the work.…”

Section: Resultsmentioning

confidence: 99%

“…The only successful method of sexing spermatozoa is quantifying sperm DNA-binding dye, followed by flow cytometry and cell sorting. The previous experiments on bovine IVF (MILLER and HUNTER, 1987;AOYAGI et al, 1988;EYESTONE and FIRST, 1989;KĄTSKA et al, 1996;CHAMBERLAND et al, 2001;KĄTSKA-KSIĄŻKIEWICZ et al, 2005) have shown that in terms of factors affecting the outcome of the procedure bull variability has appeared to be the main source of differentiation in the efficiency of this technology. When sexed sperm is used the efficiency of IVF may be affected not only by bull effect but by differentiated susceptibility of bull spermatozoa to Hoechst 3342 staining as well (SMORĄG et al, 1999).…”

Section: Introductionmentioning

confidence: 99%

In vitro production of bovine embryos using flow-cytometrically sexed sperm

et al. 2006

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The investigation aimed to compare the effect of fresh and frozen-thawed X and Y fractions of flowcytometrically sorted bovine spermatozoa on in vitro fertilization of bovine in vitro matured oocytes and subsequent blastocyst development. Sperm cells sorted in MoFloSX cytometer were used either for IVF or frozen and stored in liquid nitrogen. Immature oocytes recovered from ovaries of slaughtered animals and matured in vitro in TCM-199 containing 20% estrus cow serum and additional granulosa cells were fertilized in vitro with fresh or frozen-thawed fractions of sorted sperm. Simultaneously, control, fresh or frozen/thawed sperm was used for IVF. A total number of 2712 IVM oocytes were fertilized with sorted and control sperm of 6 bulls. Embryo cleavage rates were significantly affected by bull (P<0.0001), sperm sexing (P<0.0001) and sperm freezing (P<0.01). Blastocysts development was affected by sperm freezing (P<0.04) and sperm sexing (P<0.01). The significant differences were shown between unsorted and sorted sperm, however no differences in embryo cleavage rates and blastocysts rates were observed between X-and Y-sperm fractions, both fresh and frozen/ thawed. There were significant differences in cleavage rates among fresh, control sperm (52.7%), X fraction (26.8%) and Y fraction (24.7%). Similar differences in cleavage rates were shown for frozen/thawed control sperm (52.8%), X fraction (33.9%) and Y fraction (26.2%). The female blastocysts were frozen for further transfer, while the hatched male blastocysts were analysed by PCR revealing 76.2% accuracy. The results suggest that there were significant differences in cleavage rates and blastocyst rates due to sperm sorting in comparison to unsorted sperm and no differences between effectiveness of X and Y fractions of spermatozoa.

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Effects of two treatments on semen from different bulls on in vitro fertilization results of bovine oocytes

Cited by 32 publications

References 21 publications

Production of Piglets Derived from In Vitro-Produced Blastocysts Fertilized and Cultured in Chemically Defined Media: Effects of Theophylline, Adenosine, and Cysteine During In Vitro Fertilization1

Production of Piglets Derived from In Vitro-Produced Blastocysts Fertilized and Cultured in Chemically Defined Media: Effects of Theophylline, Adenosine, and Cysteine During In Vitro Fertilization1

In Vitro Fertilization of Bovine Oocytes in a Chemically Defined, Protein-Free Medium Varying the Bicarbonate Concentration1

In vitro production of bovine embryos using flow-cytometrically sexed sperm

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