High bovine blastocyst development in a static in vitro production system using sofaa medium supplemented with sodium citrate and myo-inositol with or without serum-proteins

Holm, Preben Bach; Booth, P.J.; Schmidt, M.; Greve, T.; Callesen, Henrik

doi:10.1016/s0093-691x(99)00162-4

Cited by 628 publications

(392 citation statements)

References 47 publications

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“…In vitro culture of embryos After IVF, presumptive zygotes were denuded from surrounding cumulus cells in HM199 and washed three times in CM, which was based on synthetic oviductal fluid aaci (Holm et al, 1999) with some modifications (Salvador et al, 2011) and with 3 mg/ml bovine serum albumin (A-8806) instead of bovine serum. Then, presumptive zygotes were in vitro cultured in groups of ,30 to 50 zygotes in 500 ml using 4-Well Nunc R plates, or in a number of five zygotes in 50 ml microdrops, depending on the experimental group (see below in Section 'Experimental design'), at 38.58C in a humidified atmosphere with 5% CO 2 and 5% O 2 in air.…”

Section: Methodsmentioning

confidence: 99%

“…For IVF, a final concentration of 1 3 10 6 sperm/ml was used. After in vitro maturation, COCs were washed three times in fertilisation Fert-Talp medium (Parrish et al, 1988) with 10 mg/ml of heparin (H9399) without epinephrine and hypotaurine and coincubated with the spermatozoa, in 500 ml using 4-Well Nunc R plates (Nunc, Fisher Scientific, Madrid, Spain), for 18 to 20 h in 5% CO 2 at 38.58C.In vitro culture of embryos After IVF, presumptive zygotes were denuded from surrounding cumulus cells in HM199 and washed three times in CM, which was based on synthetic oviductal fluid aaci (Holm et al, 1999) with some modifications (Salvador et al, 2011) and with 3 mg/ml bovine serum albumin (A-8806) instead of bovine serum. Then, presumptive zygotes were in vitro cultured in groups of ,30 to 50 zygotes in 500 ml using 4-Well Nunc R plates, or in a number of five zygotes in 50 ml microdrops, depending on the experimental group (see below in Section 'Experimental design'), at 38.58C in a humidified atmosphere with 5% CO 2 and 5% O 2 in air.…”

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confidence: 99%

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Effect of supplementation of different growth factors in embryo culture medium with a small number of bovine embryos on in vitro embryo development and quality

Ahumada

Salvador

Cebrián-Serrano

et al. 2013

Animal

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When embryos are cultured individually or in small groups, blastocyst yield efficiency and quality are usually reduced. The aim of this work was to investigate the effect of supplementation of the embryo culture medium (CM) with several growth factors (GFs) on embryo development and apoptosis rate when a reduced number of embryos were in vitro cultured. Two experimental studies (ES) were carried out. In ES 1, five treatments were tested to study the effect of GF on embryo development: Control (,30 to 50 embryos cultured in 500 ml of CM); Control 5 (Five embryos cultured in 50 ml microdrops of CM), without addition of GF in either of the two control groups; epidermal GF (EGF); IGF-I; and transforming GF-a (TGF-a) (Five embryos were cultured in 50 ml microdrops of CM with 10 ng/ml EGF, 10 ng/ml IGF-I or 10 ng/ml TGF-a, respectively). In ES 2, following the results obtained in ES 1, four different treatments were tested to study their effect on embryo development and quality (number of cells per blastocyst and apoptotic rate): Control; Control 5; EGF, all three similar to ES 1; EGF 1 IGF-I group (five embryos cultured in 50 ml microdrops of CM with 10 ng/ml EGF and 10 ng/ml IGF-I). In both ESs, it was observed that a higher proportion of embryos cultured in larger groups achieved blastocyst stage than embryos cultured in reduced groups (22.6% v. 14.0%, 12.6% and 5.3% for Control v. Control 5, IGF-I, TGF-a groups in ES 1, and 24.9% v. 17.1% and 19.0% for Control v. Control 5 and EGF in ES 2, respectively; P , 0.05), with the exception of embryos cultured in medium supplemented with EGF (18.5%) or with EGF 1 IGF-I (23.5%), in ES 1 and ES 2, respectively. With regard to blastocyst quality, embryos cultured in reduced groups and supplemented with EGF, alone or combined with IGF-I, presented lower apoptosis rates than embryos cultured in reduced groups without GF supplementation (11.6% and 10.5% v. 21.9% for EGF, EGF 1 IGF-I and Control 5 groups, respectively; P , 0.05). The experimental group did not affect the total number of cells per blastocyst. In conclusion, this study showed that supplementation of the CM with EGF and IGF could partially avoid the deleterious effect of in vitro culture of small groups of bovine embryos, increasing blastocyst rates and decreasing apoptosis rates of these blastocysts.Keywords: bovine embryo, reduced groups, in vitro culture, growth factor, apoptosis ImplicationsIn bovine, in vitro embryo production (IVEP) is especially interesting when applying the ovum pick-up technique (OPU). However, a small number of oocytes per female are usually recovered after OPU, and when embryos are cultured individually or in a reduced group blastocyst yield efficiency and quality are generally reduced. The improvement of IVEP efficiency and quality with low numbers of embryos would be of great interest for applying this technique in the dairy industry. IntroductionIn vitro embryo production (IVEP) has numerous applications in various scientific areas such as animal production or biomedicine. IVEP-rela...

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Section: Methodsmentioning

confidence: 99%

mentioning

confidence: 99%

Effect of supplementation of different growth factors in embryo culture medium with a small number of bovine embryos on in vitro embryo development and quality

Ahumada

Salvador

Cebrián-Serrano

et al. 2013

Animal

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“…Embryo culture was performed in modified synthetic oviduct fluid (SOF) containing amino-acids, citrate and myo-inositol [24] adjusted to 285 mOsm and pH 7.2-7.3. The FCS (5%) was added at 42 h post-fertilization (post-FIV).…”

Section: Embryo Culturementioning

confidence: 99%

Bovine oocyte vitrification before or after meiotic arrest: effects on ultrastructure and developmental ability

et al. 2005

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The nuclear stage at which oocytes are cryopreserved influences further development ability and cryopreservation affects ultrastructure of both cumulus cells and the oocyte. In this work, we analyze the effects of vitrification at different nuclear and cytoplasmic maturation stages on the oocyte ultrastructure and developmental ability. Culture in TCM199 + PVA with roscovitine 25 M during 24 h led to meiotic arrest (MA) in cumulus-oocyte complexes (COCs), while permissive in vitro maturation (IVM) was performed in TCM199, 10% FCS, FSH-LH and 17b-estradiol for 24 h. Oocytes were vitrified using the open pulled straw method (OPS) with minor modifications. Fresh and vitrified/warmed COCs were fixed as immature, after IVM, after meiotic arrest (MA) and after MA + IVM.Vitrification combined with MA followed by IVM produced the highest rates of degeneration, regardless of the vitrification time. As a consequence, lower proportions of embryos cleaved in these groups, although differences were eliminated at the five-eight cell stage. Development rates up to day 8 were similar in all experimental groups, being significantly lower than those in fresh controls. Only oocytes vitrified after IVM were able to give blastociysts. The morphological alterations observed can be responsible for compromised development. More research is needed to explain the low survival rates of the bovine oocyte after vitrification and warming. #

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“…In Experiment 1, up to 30 presumptive zygotes (6 hpi) were cultured in a 250-µL microdrop of modified synthetic oviduct fluid (mSOF) [28], supplemented with 30 µL/mL essential amino acids solution (× 50, Gibco-11130), 10 µL/mL non-essential amino acids solution (× 100, Gibco-11140) and 5% FBS at 39.0 ºC under 5% CO 2 , 5% O 2 and 90% N 2 for 8 d.…”

Section: In Vitro Fertilization and Culturementioning

confidence: 99%

High incidence of multiple aster formation in vitrified-warmed bovine oocytes after in vitro fertilization

Hara

Hwang

Kagawa³

et al. 2012

Theriogenology

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In vitro-matured bovine oocytes do not tolerate vitrification as well as mature murine or human oocytes. Delayed first cleavage in vitrified and in vitro-fertilized bovine oocytes may be responsible for the decreased yield of blastocysts in vitro. Since formation of sperm-aster and the subsequent assembly of microtubule network play an important role for migration and fusion of both pronuclei, aster formation in vitrified-warmed oocytes was analyzed by confocal laser-scanning microscopy. At 10 h post-insemination (hpi), proportions of oocytes fertilized normally were comparable between the vitrified and fresh control groups (67 and 70%, respectively). Proportions of oocytes that exhibited microtubule assembly were similar between the two groups (95% each), but the proportion of oocytes with multiple asters was higher in the vitrified group when compared with the fresh control group (68 vs 29%, P < 0.05). Both migration and development of two pronuclei were adversely affected by multiple aster formation. In the next experiment, multiple asters observed in 5.5 versus 8 hpi pronuclear zygotes were located near the male pronucleus, suggesting that those multiple asters were not the cytoplasmic asters of maternal origin. In conclusion, multiple aster formation frequently observed in vitrified-warmed bovine oocytes may be related to loss of ooplasmic function responsible for normal microtubule assembly from the sperm-aster.

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High bovine blastocyst development in a static in vitro production system using sofaa medium supplemented with sodium citrate and myo-inositol with or without serum-proteins

Cited by 628 publications

References 47 publications

Effect of supplementation of different growth factors in embryo culture medium with a small number of bovine embryos on in vitro embryo development and quality

Effect of supplementation of different growth factors in embryo culture medium with a small number of bovine embryos on in vitro embryo development and quality

Bovine oocyte vitrification before or after meiotic arrest: effects on ultrastructure and developmental ability

High incidence of multiple aster formation in vitrified-warmed bovine oocytes after in vitro fertilization

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