Effect of bovine sperm separation by either swim-up or Percoll method on success of in vitro fertilization and early embryonic development

Parrish, J.J.; Krogenæs, Anette; Susko-Parrish, J.L.

doi:10.1016/0093-691x(95)00271-9

Cited by 370 publications

(252 citation statements)

References 28 publications

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“…In bovine, the embryo culture conditions are a well documented process (Parrish et al, 1994;Holm et al, 1999;Papis et al, 2007;Sargirkaya et al, 2007;Adona et al, 2008;Corrêa et al, 2008). Oocyte in-vitro maturation is a viable phenomenon as oocyte matured, fertilized and cultured in vitro can generate embryos with full developmental potential after embryo transfer.…”

Section: Resultsmentioning

confidence: 99%

“…For in vitro fertilization (IVF), frozen-thawed semen from the same bull was prepared according to the discontinuous Percoll gradient technique (Parrish et al, 1994). The separated sperm was rinsed twice in TALP medium and then resuspended to a final concentration of 1 × 10 6 sperm cells mL -1 in IVF medium.…”

Section: In Vitro Maturation Fertilization and Embryo Culture Procedmentioning

confidence: 99%

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Melatonin in maturation media fails to improve oocyte maturation, embryo development rates and DNA damage of bovine embryos

Takada

Martins

Mingoti

et al. 2010

Sci. agric. (Piracicaba, Braz.)

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Melatonin (MEL) acts as a powerful scavenger of free radicals and direct gonadal responses to melatonin have been reported in the literature. Few studies, however, have evaluated the effect of MEL during in vitro maturation (IVM) on bovine embryos. This study tested the addition of MEL to maturation medium (MM) with no gonadotropins on nuclear maturation and embryo development rates and the incidence of DNA damage in resulting embryos. Cumulus-oocyte complexes were aspirated from abattoir ovaries and cultured in MM (TCM-199 medium supplemented with 10% fetal calf serum -FCS) at 39°C and 5% CO 2 in air. After 24 hours of culture in MM with 0.5 μg mL -1 FSH and 5.0 μg mL -1 LH; 10 -9 M MEL) or 10 -9 M MEL, 0.5 μg mL -1 FSH and 5.0 μg mL -1 LH, the oocytes were stained with Hoechst 33342 to evaluate nuclear maturation rate. After in vitro fertilization and embryo culture, development rates were evaluated and the blastocysts were assessed for DNA damage by Comet assay. There was no effect of melatonin added to the MM, alone or in combination with gonadotropins, on nuclear maturation, cleavage and blastocyst rates. These rates ranged between 88% to 90%, 85% to 88% and 42% to 46%, respectively. The extent of DNA damage in embryos was also not affected by MEL supplementation during IVM. The addition of 10 -9 M MEL to the MM failed to improve nuclear maturation and embryo development rates and the incidence of DNA damage in resulting embryos, but was able to properly substitute for gonadotropins during IVM. Key words: comet assay, embryos, gonadotropins, in vitro fertilization, melatonin Melatonina no meio de maturação não melhorou as taxas de maturação dos ovócitos, de desenvolvimento embrionário e a fragmentação do DNA dos embriões bovinos RESUMO: Melatonin (MEL) atua como um potente redutor de radicais livres. Efeito direto da MEL na função gonadal também foi observado. Existem poucos estudos relacionados ao efeito da MEL durante a maturação no desenvolvimento embrionário in vitro. Avaliou-se a adição de MEL no meio de maturação (sem gonadotrofinas) nas taxas de maturação nuclear e de desenvolvimento embrionário e na incidência de fragmentação do DNA nos embriões produzidos in vitro. Complexos cumulus-ovócitos foram aspirados de ovários provenientes de matadouros e cultivados em meio de maturação (Meio TCM-199 suplementado com 10% de soro fetal bovino) a 39°C e 5% CO 2 em ar. Após 24 horas de cultivo em meio de maturação com 0,5 μg mL -1 de FSH e 5,0 μg mL -1 de LH; 10 -9 M de MEL ou 10 -9 M de MEL, 0,5 μg mL -1 de FSH e 5,0 μg mL -1 de LH, os ovócitos foram corados com Hoechst 33342 para avaliação da taxa de maturação nuclear. A fragmentação do DNA dos blastocistos foi avaliada pela técnica do Cometa. Não houve efeito da MEL adicionada ao meio de maturação, com ou sem gonadotrofinas, nas taxas de maturação nuclear, clivagem e blastocistos. Os percentuais variaram entre 88% a 90%, 85% a 88% e 42% a 46%, respectivamente. O grau de fragmentação do DNA também não foi afetado pela adição de MEL ao meio de matur...

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Section: Resultsmentioning

confidence: 99%

Section: In Vitro Maturation Fertilization and Embryo Culture Procedmentioning

confidence: 99%

Melatonin in maturation media fails to improve oocyte maturation, embryo development rates and DNA damage of bovine embryos

Takada

Martins

Mingoti

et al. 2010

Sci. agric. (Piracicaba, Braz.)

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“…Motile spermatozoa were obtained by centrifugation of the pellet at 700 g for 30 min on a discontinuous gradient of Percoll (Amersham Pharmacia Biotech; 2 ml 45% in BTS over 2 ml 90%). The 90% Percoll solution contained 80 mM NaCl, 10 mM HEPES, 3.1 mM KCl, 0.3 mM NaH 2 PO 4 , 25 mM NaHCO 3 , 0.4 mM MgCl 2 , 2 mM CaCl 2 , and 21.6 mM lactic acid (Eid et al 1994, Parrish et al 1995. Sperm cells collected at the bottom of the 90% fraction were washed by centrifugation at 100 g for 10 min in 4 ml modified Trisbuffered medium (mTBM; Marchal et al 2003).…”

Section: Preparation Of Equine and Porcine Spermatozoamentioning

confidence: 99%

Deleted in malignant brain tumor 1 is secreted in the oviduct and involved in the mechanism of fertilization in equine and porcine species

Ambruosi¹,

Accogli²,

Douet³

et al. 2013

Reproduction

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Oviductal environment affects preparation of gametes for fertilization, fertilization itself, and subsequent embryonic development. The aim of this study was to evaluate the effect of oviductal fluid and the possible involvement of deleted in malignant brain tumor 1 (DMBT1) on IVF in porcine and equine species that represent divergent IVF models. We first performed IVF after pre-incubation of oocytes with or without oviductal fluid supplemented or not with antibodies directed against DMBT1. We showed that oviductal fluid induces an increase in the monospermic fertilization rate and that this effect is canceled by the addition of antibodies, in both porcine and equine species. Moreover, pre-incubation of oocytes with recombinant DMBT1 induces an increase in the monospermic fertilization rate in the pig, confirming an involvement of DMBT1 in the fertilization process. The presence of DMBT1 in the oviduct at different stages of the estrus cycle was shown by western blot and confirmed by immunohistochemical analysis of ampulla and isthmus regions. The presence of DMBT1 in cumulus-oocyte complexes was shown by western blot analysis, and the localization of DMBT1 in the zona pellucida and cytoplasm of equine and porcine oocytes was observed using immunofluorescence analysis and confocal microscopy. Moreover, we showed an interaction between DMBT1 and porcine spermatozoa using surface plasmon resonance studies. Finally, a bioinformatic and phylogenetic analysis allowed us to identify the DMBT1 protein as well as a DMBT1-like protein in several mammals. Our results strongly suggest an important role of DMBT1 in the process of fertilization.

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“…After allowing 18 h for fertilisation, the oocyte or presumptive zygote was transferred into a drop of development medium. The development medium was Synthetic Oviductal fluid (SOF, EmbryoMax; Chemicon International) and was supplemented with fatty acid free BSA (8 mg/ml), pyruvate (0.2 mM), BME (essential amino acids, 50!, 10 ml/ml), MEM (non-essential amino acids, 100!, 20 ml/ml), and gentamicin (50 mg/ml) as described previously (Parrish et al 1995). The oocyte was examined for indications of cleavage under a stereomicroscope 48 h after exposure to spermatozoa.…”

Section: Ivf and Embryo Developmentmentioning

confidence: 99%

Relationship of vascular perfusion of the wall of the preovulatory follicle to in vitro fertilisation and embryo development in heifers

Siddiqui¹,

Gastal²,

Gastal³

et al. 2009

Reproduction

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The effect of the extent of vascular perfusion of the wall of the preovulatory follicle on in vitro cleavage rate of the recovered oocyte and embryo development to >8 cells was studied in 52 heifers. Heifers received a luteolytic dose of prostaglandin F2α (PGF2α) when the largest follicle was ≥11 mm. An ovulation-inducing injection of GnRH was given 36 h later (hour 0), and collection of follicular fluid and the oocyte was done at hour 26. Vascular perfusion of the follicular wall was assessed by colour Doppler ultrasonography at hours 0 and 26. Each of the recovered oocytes (41/52; 79%) was mature (extruded polar body). Cleavage and embryo development were assessed at 48 h and 120 h respectively, after in vitro fertilisation (IVF). The percentage of cleaved oocytes and >8 cell embryos was 80% (31/39) and 55% (17/31) respectively. Vascular perfusion of the follicular wall was greater (lower pulsatility index; P<0.001) for follicles that produced cleaved versus non-cleaved oocytes and greater (P<0.04) for follicles that produced >8 cell versus ≤8 cell embryos. Percentage of follicular wall with Doppler signals of blood flow was greater (P<0.001) for >8 cell versus ≤8 cell embryos. Follicular-fluid concentration of free IGF1 was lower for cleaved oocytes (P<0.001) and >8 cell embryos (P<0.05), and oestradiol was lower (P<0.05) for >8 cell embryos. Results supported the hypothesis that greater vascular perfusion of the wall of the preovulatory follicle was positively associated with IVF and embryo development.

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Effect of bovine sperm separation by either swim-up or Percoll method on success of in vitro fertilization and early embryonic development

Cited by 370 publications

References 28 publications

Melatonin in maturation media fails to improve oocyte maturation, embryo development rates and DNA damage of bovine embryos

Melatonin in maturation media fails to improve oocyte maturation, embryo development rates and DNA damage of bovine embryos

Deleted in malignant brain tumor 1 is secreted in the oviduct and involved in the mechanism of fertilization in equine and porcine species

Relationship of vascular perfusion of the wall of the preovulatory follicle to in vitro fertilisation and embryo development in heifers

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