Addition of penicillamine, hypotaurine and epinephrine (PHE) or bovine oviductal epithelial cells (BOEC) alone or in combination to bovine in vitro fertilization medium increases the subsequent embryo cleavage rate

Miller, Gad; Gliedt, D.W.; Rakes, J. M.; Rorie, R.W.

doi:10.1016/0093-691x(94)90178-l

Cited by 32 publications

(17 citation statements)

References 27 publications

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“…Our findings comes in the same line with Miller et al (1994) and Goncalves et al (2014) who reported that PHE supplementation improves the cleavage rate, increases fertilization and blastocyst formation rates in cattle, respectively.…”

Section: Resultssupporting

confidence: 82%

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Improvement of in Vitro Fertilization in Buffalo by Increasing the Fertilizing Capacity of Spermatozoa

ElRuby¹,

Hussein²,

Bedier³

et al. 2017

AJVS

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Key words: D-Penecillamine, Hypotaurine, Epinephrine, In vitro fertilization, BuffaloIn vitro fertilization (IVF) is a necessary procedure in the in vitro production of embryo. Spermatozoa need to undergo capacitation to be able to perform the acrosome reaction which is required for zona pellucida penetration. Different additives are used to aid in that process, such as caffeine and theophylline. In this study, the effects of IVF media supplementation with D-penicillamine, Hypotaurine and epinephrine (PHE) on fertilization and cleavage rates were assessed. Cumulus oocyte complexes (COCs) containing a compact cumulus cells and normal homogenous cytoplasm were selected. In vitro maturation (IVM) was performed in TCM199 supplied with hormones overnight in a culture conditions of 38.5°C and 5% CO 2 in air. After 20-22 hr. of maturation, oocytes were randomly assigned to five groups of sperm-TALP media (control and four different PHE concentrations) then inseminated. The fertilization rate of the expected embryos were assessed after 22 hr. after staining with aceto-orcin 1% or cultured for 48 hr. to estimate the cleavage rate. The current data revealed that, 25 µl/ml PHE significantly (P< 0.05) improved fertilization (55.93 vs 36.96%) and cleavage (46.15 vs 22.22%) rates when compared to control.

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Section: Resultssupporting

confidence: 82%

“…PHE stock was prepared according to Miller et al (1994). Sperm-TALP media was prepared according to Parrish (2014) with 20µg/ml heparin sulfate addition to the media before filtration, The pH was adjusted to 7.6.…”

Section: Methodsmentioning

confidence: 99%

Improvement of in Vitro Fertilization in Buffalo by Increasing the Fertilizing Capacity of Spermatozoa

ElRuby¹,

Hussein²,

Bedier³

et al. 2017

AJVS

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“…In a number of studies, P was found to enable an increase in the percentage of spermatozoa undergoing the acrosome reaction in the presence of epinephrine [49][50][51]. However, these experiments were carried out in undefined conditions, and the efficacy of being able to induce acrosome reactions was attributed to epinephrine or hypotaurine, without regard for P. Including 0.5 mg/ml P + PVA has supported proportions of morula, blastocyst, and expanded blastocyst development in vitro similar to the those supported by undefined conditions (experiment 1).…”

Section: Discussionmentioning

confidence: 99%

In Vitro Developmental Competence of in Vitro-Matured Bovine Oocytes Fertilized and Cultured in Completely Defined Media1

Keskintepe

Brackett

1996

Biology of Reproduction

155

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The objective was to establish an in vitro system in which bovine oocytes can be matured, fertilized, and cultured up to the blastocyst stage without support of serum, BSA, or somatic cells. Media consisted of modified tissue culture medium 199 (mTCM 199) with ovine LH (oLH) for maturation (IVM), experimental alterations of modified defined medium (mDM) for sperm selection and insemination (IVF), and citrate+synthetic oviductal fluid+nonessential amino acids (c-SOF+NEA) for zygote/embryo culture (IVC). Effects of heparin, BSA, polyvinyl alcohol (PVA), penicillamine (P), Hepes, and sodium bicarbonate (NaHCO3) were studied. Results included proportions of oocytes that cleaved by 48 h and that reached morulae by 120 h, blastocysts by 168 h, and expanded blastocysts by 216 h postinsemination (pi). Best results were obtained when the IVF medium included 0.5 mg P+1.0 mg PVA per milliliter with no more than 10 mM Hepes, and when 3.0 mg PVA/ml and 10 mM Hepes were present for IVC. Different concentrations of NaHCO3, up to 50 mM from 25 mM, during IVF did not alter results. Embryos produced in defined conditions yielding the best results remained viable after vitrification as evidenced by continued development in vitro for 96 h postthawing. Bovine oocytes matured in defined medium supplemented with LH were fertilized and cultured up to the blastocyst stage in chemically defined conditions that afforded results comparable to those reported earlier after inclusion of serum, BSA, and/or somatic cells.

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“…As agglutination occurs when trying to capacitate ram spermatozoa in TALP, it was important to assess if the inclusion of PEN in this medium would interfere in the capacitation process. It would seem unlikely as low doses of PEN (20 mM), in conjunction with hypotaurine and epinephrine (PHE), have been included in multiple bovine IVF protocols to improve oocyte penetration and embryo quality (Miller et al 1994, Goncalves et al 2014, Kang et al 2015. PEN has also been used in protein-free media to promote capacitation (Andrews & Bavister 1989, Keskintepe & Brackett 1996.…”

Section: Discussionmentioning

confidence: 99%

Penicillamine prevents ram sperm agglutination in media that support capacitation

Leahy¹,

Rickard²,

Aitken³

et al. 2016

Reproduction

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Ram spermatozoa are difficult to capacitate in vitro. Here we describe a further complication, the unreported phenomenon of headto-head agglutination of ram spermatozoa following dilution in the capacitation medium Tyrodes plus albumin, lactate and pyruvate (TALP). Sperm agglutination is immediate, specific and persistent and is not associated with a loss of motility. Agglutination impedes in vitro sperm handling and analysis. So the objectives of this study were to investigate the cause of sperm agglutination and potential agents which may reduce agglutination. The percentage of non-agglutinated, motile spermatozoa increased when bicarbonate was omitted from complete TALP suggesting that bicarbonate ions stimulate the agglutination process. D-penicillamine (PEN), a nucleophilic thiol, was highly effective at reducing agglutination. The inclusion of 250 mM PEN in TALP reduced the incidence of motile, agglutinated spermatozoa from 76.7G2.7% to 2.8G1.4%. It was then assessed if PEN (1 mM) could be included in existing ram sperm capacitation protocols (TALP C1 mM dibutyryl cAMP, caffeine and theophylline) to produce spermatozoa that were simultaneously capacitated and non-agglutinated. This protocol resulted in a sperm population which displayed high levels of tyrosine phosphorylated proteins and lipid disordered membranes (merocyanine-540) while remaining motile, viable, acrosome-intact and non-agglutinated. In summary, PEN (1 mM) can be included in ram sperm capacitation protocols to reduce sperm agglutination and allow for the in vitro assessment of ram sperm capacitation.

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Addition of penicillamine, hypotaurine and epinephrine (PHE) or bovine oviductal epithelial cells (BOEC) alone or in combination to bovine in vitro fertilization medium increases the subsequent embryo cleavage rate

Cited by 32 publications

References 27 publications

Improvement of in Vitro Fertilization in Buffalo by Increasing the Fertilizing Capacity of Spermatozoa

Improvement of in Vitro Fertilization in Buffalo by Increasing the Fertilizing Capacity of Spermatozoa

In Vitro Developmental Competence of in Vitro-Matured Bovine Oocytes Fertilized and Cultured in Completely Defined Media1

Penicillamine prevents ram sperm agglutination in media that support capacitation

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