Male-dependent variability of fertilization and embryo development in two bovine in vitro fertilization systems and the effects of casein phosphopeptides (CPPs)

1997 ◽  
Vol 9 (4) ◽  
pp. 465 ◽  
Author(s):  
U. Kreysing ◽  
T. Nagai ◽  
H. Niemann
Keyword(s):  
In Vitro Fertilization ◽  
Embryo Development ◽  
Individual Variability ◽  
Bovine Embryo ◽  
Blastocyst Formation ◽  
Embryo Production ◽  
Vitro Fertilization ◽  
Casein Phosphopeptides ◽  
Fertilization System

This study investigated the effects of semen from five different bulls and two different ejaculates of the same bull on penetration, cleavage, blastocyst formation, and cell allocation in bovine blastocysts produced in vitro. Casein phosphopeptides (CPPs) were tested for their ability to enhance fertilization and minimize variability among bulls and ejaculates. In Experiment 1, the BO-fertilization system was employed. Penetration and polyspermy both displayed great variation among bulls and between ejaculates, whereas no significant differences were observed in cleavage and blastocyst-formation rates. Similar variability was observed in penetration, polyspermy, cleavage, blastocyst-formation rates and cell allocation and distribution when the two fertilization systems, TALP and BO, were compared in Experiment 2. The BO-system supported penetration and polyspermy better (P < 0·05) than the TALP-system, whereas the TALP-system was superior (P < 0·05) in supporting cleavage and blastocyst formation. Significant interactions existed between bulls and the fertilization system employed. It is concluded that the success of in vitrofertilization is markedly dependent on individual bulls as well as on ejaculates from the same bull. CPPs are able to enhance penetration and embryo development in certain bulls or ejaculates and thus contribute to reducing the degree of individual variability, but they do not generally improve the success of bovine embryo production in vitro.

238 KNOCKING DOWN OF THYROID HORMONE RECEPTORS INHIBITS DEVELOPMENT OF EARLY BOVINE EMBRYOS

2013 ◽  
Vol 25 (1) ◽  
pp. 267
Author(s):  
N. Y. Rho ◽  
F. A. Ashkar ◽  
T. Revay ◽  
P. Madan ◽  
W. A. King
Keyword(s):  
Embryo Development ◽  
Reference Genes ◽  
Messenger Rna ◽  
Bovine Embryo ◽  
Bovine Embryos ◽  
Blastocyst Formation ◽  
Embryo Production ◽  
Mrna Transcripts ◽  
In Vitro Embryo Production

Thyroid hormones (TH) play an important role in the physiology of vertebrates, ranging from the regulation of metabolic processes to cell proliferation, differentiation, and embryo development. We have previously shown a beneficial effect of supplementing TH in in vitro embryo production media. Recently, detection of TH receptors (TR) in oocytes and early stages of pre-implantation embryos indicated a possible regulatory role for TH in these stages (unpublished data). The objective of this study was to investigate the importance of TR expression in the pre-attachment bovine embryo in vitro. Bovine embryos, produced by standard in vitro embryo production procedures, were microinjected at the zygote stage with small interfering RNA (siRNA) specifically designed for knocking down either TR-α or TR-β. In addition, groups of zygotes were microinjected with scrambled siRNA (SI) or were not injected (NI), and these groups served as controls. Embryo developmental rates were assessed using light microscopy for blastocyst formation rates and expression of TR messenger RNA (mRNA) transcripts at the blastocyst stage was assessed by quantitative PCR across all groups. Expression of TR mRNA was normalized against glyceraldehyde 3-phosphate dehydrogenase, H2a, and 18S as reference genes. There was a significant decrease in blastocyst formation rates in both embryo groups injected with either TR-α (P < 0.002) and TR-β (P < 0.001) siRNA compared with the NI and SI groups. Moreover, the TR-β knockdown group exhibited a lower developmental rate than the TR-α knockdown group, which indicates a stronger inhibitory role for TR-β. Quantification of the level of TR mRNA expression in four groups normalized with three different reference genes shows a consistent significant reduction in the levels of TR-α (P < 0.05) and TR-β (P < 0.02) mRNA transcripts compared with the NI and SI groups. However, TR-β expression was inhibited more than was TR-α expression. In conclusion, the results indicate that knocking down either TR-α or TR-β restrains embryo development. This suggests that TH play a vital role in the regulation of embryo development through their receptors during bovine early embryogenesis. The specific role of each of these receptors and their mechanism of action in mediating development needs to be further elucidated. Funding was provided by CRC, NSERC, and the EmbryoGENE network.

scholarly journals Evaluation of in vitro production rates of bovine embryos using melatonin-supplemented culture medium

2021 ◽  
Vol 10 (6) ◽  
pp. e19010615544
Author(s):  
Ricardo Magalhães ◽  
Carlos Renato de Freitas Guaitolini ◽  
Marcio Luiz Denck Tramontin ◽  
Danielle Andressa Oliveira Sestari ◽  
Bruno Argenton de Barros ◽  
...  
Keyword(s):  
Culture Medium ◽  
Statistical Significance ◽  
Gradient Centrifugation ◽  
Control Group ◽  
Bovine Embryo ◽  
Blastocyst Formation ◽  
In Vitro Production ◽  
Embryo Production ◽  
Vitro Fertilization

In this study, we aimed to evaluate the rate of bovine embryo production by using 50 ng/mL melatonin supplementation in in vitro culture medium. For this, oocytes from slaughterhouse ovaries were matured in vitro in TCM-199 medium with Earle’s balanced salt solution + 10% SFB, FSH, and LH in an atmosphere of 5% CO2. Twenty-four hours after IVM, the oocytes underwent in vitro fertilization in human tubal fluid under the same conditions as above, for 18 h. Semen was fractionated by Percoll gradient centrifugation and the concentration of sperm was adjusted to 1 × 106/mL. Probable zygotes were then divided into two groups: the control group grown in drops of 90 μL SOFaa medium + 0.6% BSA + 2.5% SFB, in an atmosphere of 5% CO2, 90% N2, and a melatonin group (Mel), similarly cultured in 90 μL drops of SOFaa medium + 0.6% BSA + 2.5% SFB + 50 ng/mL melatonin. Cleavage rates were assessed on day 3 (D3). On D7, blastocyst formation rates were evaluated. Eight routines were performed (320 oocytes per routine). Data were analyzed with ANOVA, followed by Tukey’s range test using a general linear model. The level of statistical significance was set at 5%. There were no differences in the rates of cleavage or blastocyst formation between the control and melatonin groups (P > 0.05). Thus, under the conditions used in this study, supplementation with melatonin did not yield benefits in increasing the rate of in vitro bovine embryo production.

177 EFFECT OF DIFFERENT FERTILIZATION MEDIA ON IN VITRO BOVINE EMBRYO DEVELOPMENT USING FLOW-CYTOMETRICALLY-SORTED FEMALE SPERM

2014 ◽  
Vol 26 (1) ◽  
pp. 203
Author(s):  
L. B. Ferré ◽  
Y. S. Bogliotti ◽  
J. L. Chitwood ◽  
P. J. Ross
Keyword(s):  
In Vitro Fertilization ◽  
Embryo Development ◽  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Cumulus Cell ◽  
Bovine Embryo ◽  
Vitro Fertilization ◽  
Female Sex ◽  
Fetal Bovine

High demand exists for in vitro-derived bovine embryos fertilized with female sex-sorted sperm by seedstock and commercial cattle producers. The aim of this study was to evaluate different fertilization media on in vitro fertilization performance using female sex-sorted semen. Ovaries were collected from a slaughterhouse and oocytes aspirated from 2- to 6-mm follicles. Cumulus-oocyte complexes containing compact and complete cumulus cell layers were selected and matured in groups of 50 cumulus-oocyte complexes in 400 μL of M199 medium supplemented with ALA-glutamine (0.1 mM), Na pyruvate (0.2 mM), gentamicin (5 μg mL–1), epidermal growth factor (50 ng mL–1), oFSH (50 ng mL–1), bLH (3 μg mL–1), cysteamine (0.1 mM), and 10% fetal bovine serum for 22 to 24 h. Fertilization (Day 0) was carried out using female sex-sorted semen selected with a discontinuous density gradient and diluted to a final concentration of 1 × 106 sperm mL–1. Three different fertilization media, M199 (Gibco 11043–023, Grand Island, NY, USA), SOF (Tervit et al. 1972 J. Reprod. Fertil. 30, 493–497), and TALP (Parrish et al. 1988 Biol. Reprod. 38, 1171–1180), were assayed along with 3 female sex-sorted bulls. All fertilization media were supplemented with fructose (90 μg mL–1), penicillamine (3 μg mL–1), hypotaurine (11 μg mL–1), and heparin (20 μg mL–1). After 18 h, presumptive zygotes were denuded and cultured in groups of 15 to 20 in 50-μL drops of KSOM-BSA for 9 days. On Day 3, 3% fetal bovine serum was added. Low oxygen tension (5% O2) was used for the entire culture period. On Days 7 and 9 blastocysts and hatched embryos, respectively, were morphologically evaluated according to IETS standards and recorded. Results are shown in Table 1. Data was compared by chi-squared analysis. Fertilization media affected cleavage rate and subsequent embryo development, quality, and hatching ability. The SOF and TALP fertilization media produced significantly more and higher quality embryos than M199. Table 1.In vitro fertilization performance after oocyte fertilization using sex-sorted sperm

Impact of in vitro fertilization of bovine oocytes with sex-sorted frozen–thawed spermatozoa on developmental kinetics, quality and sex ratio of developing embryos

Zygote ◽  
2010 ◽  
Vol 18 (3) ◽  
pp. 185-194 ◽  
Author(s):  
J. Peippo ◽  
M. Räty ◽  
K. Korhonen ◽  
M. Eronen ◽  
K. Kananen ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Sex Ratio ◽  
Embryo Development ◽  
Bovine Embryos ◽  
Embryo Production ◽  
Male Embryo ◽  
Vitro Fertilization ◽  
Bovine Oocytes

SummaryWe studied whether bovine embryos developing after in vitro fertilization (IVF) with sex-sorted spermatozoa differed in developmental kinetics, quality and sex ratio from embryos produced with unsorted spermatozoa. Abattoir-derived oocytes were fertilized with X-sorted, Y-sorted or unsorted spermatozoa from a single bull. To evaluate economical use of the sex-sorted spermatozoa, washed spermatozoa from a single straw (2 million spermatozoa) were used to fertilize each batch of collected oocytes without any further isolation steps. Concentration of the unsorted spermatozoa was adjusted accordingly. Fertilizations were assessed by staining sperm asters at 10 hpi and pronuclei at 20 hpi. Embryo development and morphological quality were monitored on days 2, 7, 8 and 9 of the development (IVF = day 0). All embryos were sexed using PCR. Following fertilization, penetration and subsequent cleavage rates were compromised in the X-sorted group compared with the Y-sorted and unsorted groups (penetration: 58.0% vs. 89.8% and 90.0%, cleavage: 65.3% vs. 81.5% and 75.0%). The use of the sex-sorted spermatozoa did not, however, reduce the proportion of transferable embryos (sex-sorted 29.6% vs. unsorted 27.7%) or their quality (quality 1: sex-sorted 36.0% vs. unsorted 19.9%). The Y-sorted spermatozoa produced more transferable embryos of better quality than the X-sorted spermatozoa (days 7–8: 31.9% vs. 26.4%, quality 1: 38.9% vs. 30.6%). On average, out of 10 transferable embryos, nine were of the predicted sex in the X- and Y-sorted spermatozoa groups. These results indicate that low numbers of X- and Y-sorted spermatozoa can be used successfully for female and male embryo production in vitro.

Effect of nicotinamide supplementation in in vitro fertilization medium on bovine embryo development

2020 ◽  
Vol 87 (10) ◽  
pp. 1070-1081
Author(s):  
Yu‐Guo Yuan ◽  
Ayman Mesalam ◽  
Seok‐Hwan Song ◽  
Kyeong‐Lim Lee ◽  
Lianguang Xu ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Embryo Development ◽  
Bovine Embryo ◽  
Vitro Fertilization

Exogenous glutathione supplementation in culture medium improves the bovine embryo development after in vitro fertilization

2015 ◽  
Vol 84 (5) ◽  
pp. 716-723 ◽  
Author(s):  
Wei-Jun Sun ◽  
Yun-Wei Pang ◽  
Yan Liu ◽  
Hai-Sheng Hao ◽  
Xue-Ming Zhao ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Embryo Development ◽  
Culture Medium ◽  
Bovine Embryo ◽  
Vitro Fertilization
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