219 IN VITRO FERTILIZATION RATE OF MATURED PIG OOCYTES BY FROZEN - THAWED KOLBROEK PIG SPERM CELLS

2011 ◽  
Vol 23 (1) ◽  
pp. 208
Author(s):  
M. H. Mapeka ◽  
K. C. Lehloenya ◽  
M. L. Mphaphathi ◽  
T. L. Nedambale
Keyword(s):  
In Vitro Fertilization ◽  
South African ◽  
Fetal Bovine Serum ◽  
Fertilization Rate ◽  
Blastocyst Stage ◽  
Sperm Cells ◽  
Department Of Agriculture ◽  
Vitro Fertilization ◽  
Fetal Bovine

No studies have investigated the IVF rate of South African indigenous Kolbroek sperm cells following cryopreservation. The objective of this study was to test if frozen–thawed Kolbroek pig sperm cells could penetrate pig oocytes matured in vitro. Pig ovaries were collected from a local abattoir and cumulus–oocytes complexes were obtained by aspiration and were then in vitro matured in TCM-199 supplemented with 10% pig follicular fluid, 10% fetal bovine serum, and 1 μg mL–1 of FSH and LH. Following 44 h of incubation, 200 matured pig oocytes were randomly assigned to 2 treatments with frozen–thawed and fresh (control) Kolbroek pig sperm cells. For IVF, Kolbroek sperm cells were in vitro capacitated using Brackett and Oliphant’s sperm wash medium. Matured pig oocytes and sperm cells were co-incubated for 24 h in Brackett and Oliphant’s IVF medium. Following fertilization, presumptive zygotes were in vitro cultured at 39°C in 5% CO2, 5% O2, and 90% N2. Rate of fertilization was identified by the number of cleaved zygotes. Data were analysed by ANOVA. The total motility of Kolboek pig sperm cells used for IVF was 40% for frozen–thawed sperm cells and 80% for fresh sperm cells. The results showed that Kolbroek pig sperm cells were able to penetrate pig oocytes in vitro. However, no significant (P < 0.05) difference was observed in the percentage of cleavage of pig oocytes fertilized with either frozen–thawed (13.25%) or fresh (13.0%) Kolbroek pig sperm cells. The percentage of embryos that developed to the morulae stage was 2% in frozen–thawed sperm cells and was 0% in fresh Kolbroek sperm cells. Furthermore, oocytes fertilized with Kolboek sperm cells did not develop to the blastocyst stage in either treatment. In conclusion, this study demonstrated that frozen–thawed Kolbroek sperm cells are able to fertilize matured pig oocytes in vitro. This study was funded by the Department of Agriculture Forestry and Fishery, ARC, DST-PDP (RT19000), and National Research Foundation (NRF, Grant No. RT21 and 24000).

scholarly journals Effect of Bali cattle ovarian status on oocytes nuclear maturation and in vitro fertilization rate

2018 ◽  
Vol 22 (4) ◽  
pp. 173 ◽  
Author(s):  
Herry Sonjaya ◽  
M. Yusuf ◽  
A. Hamdana ◽  
Renny Fatmyah Utamy ◽  
Sri Gustina ◽  
...  
Keyword(s):  
Bovine Serum ◽  
Fetal Bovine Serum ◽  
Fertilization Rate ◽  
Reproductive Status ◽  
Vitro Fertilization ◽  
Nuclear Maturation ◽  
Fetal Bovine ◽  
Bali Cattle ◽  
Pronuclear Formation

<p>The aim of this study was to investigate whether the reproductive status influences the nuclear maturation and fertilization rates of bali cattle oocytes in vitro. Several pairs of ovary were classified into four groups: 1) ovaries with Corpus Luteum (CL) and Dominant Follicle (DF), 2) ovaries without CL and with DF, 3) ovaries with CL and without DF, 4) ovaries without both CL and DF. In the first experiment, oocytes were collected by slicing method in Phosphate Buffer Saline (PBS) medium supplemented with 10% Fetal Bovine Serum (FBS) and 100 IU/ml penicillin streptomycin. Oocytes were matured in tissue culture medium (TCM)-199 supplemented with 10% Fetal Bovine Serum (FBS), 10 IU/ml Follicle Stimulating Hormone (FSH), 10 IU/ml Luteinizing Hormone (LH), and 50 μg/ml gentamycin. Oocytes were matured in 5% CO2 incubator, 38oC for 24 h. In the second experiment, oocytes were matured and then fertilized in vitro to observe pronuclear formation. The first experiment showed that the percentage of oocytes reached methaphase-II (MII) stage on ovaries with CL and without DF (89.47%) were higher (P&lt;0,01) compared to ovaries without both CL and DF (75,47%), ovaries without CL and with DF (74.,41%), or ovaries with CL and DF (65,52%). The result of second experiment showed that the ovarian reproductive status was not significantly different (P&gt;0.05) on fertilization rate.</p>

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

scholarly journals A Brief Incubation of Cumulus-Enclosed Mouse Eggs in a Calcium-Free Medium Containing a High Concentration of Calcium-Chelator Markedly Improves Preimplantation Development

2020 ◽  
Vol 17 (10) ◽  
pp. 3505
Author(s):  
Valeria Merico ◽  
Silvia Garagna ◽  
Maurizio Zuccotti
Keyword(s):  
In Vitro Fertilization ◽  
Mammalian Species ◽  
Developmental Rate ◽  
Cumulus Cells ◽  
Fertilization Rate ◽  
Preimplantation Development ◽  
High Concentration ◽  
Vitro Fertilization ◽  
Positive Effects

The presence of cumulus cells (CCs) surrounding ovulated eggs is beneficial to in vitro fertilization and preimplantation development outcomes in several mammalian species. In the mouse, this contribution has a negligible effect on the fertilization rate; however, it is not yet clear whether it has positive effects on preimplantation development. Here, we compared the rates of in vitro fertilization and preimplantation development of ovulated B6C3F1 CC-enclosed vs. CC-free eggs, the latter obtained either after a 5 min treatment in M2 medium containing hyaluronidase or after 5–25 min in M2 medium supplemented with 34.2 mM EDTA (M2-EDTA). We found that, although the maintenance of CCs around ovulated eggs does not increment their developmental rate to blastocyst, the quality of the latter is significantly enhanced. Most importantly, for the first time, we describe a further quantitative and qualitative improvement, on preimplantation development, when CC-enclosed eggs are isolated from the oviducts in M2-EDTA and left in this medium for a total of 5 min prior to sperm insemination. Altogether, our results establish an important advancement in mouse IVF procedures that would be now interesting to test on other mammalian species.

scholarly journals Efficiency Comparison Between Dry And Humid Cultures For Clinical Outcome of The In Vitro Fertilization-Embryo Transfer

2021 ◽  
Author(s):  
Weihai Xu ◽  
Lin Zhang ◽  
Ling Zhang ◽  
Shishi Li ◽  
Jing Shu
Keyword(s):  
In Vitro Fertilization ◽  
Embryo Transfer ◽  
Implantation Rate ◽  
Fertilization Rate ◽  
Cleavage Rate ◽  
High Quality ◽  
Vitro Fertilization ◽  
Efficiency Comparison ◽  
Ivf Et

Abstract Background: In this study, we compared the in vitro embryo development, embryo transfer outcome and the offspring outcome in the in vitro fertilization-embryo transfer (IVF-ET) between dry culture (DC) and humid culture (HC). Methods: Our study was divided into two parts. Firstly, we determined the fertilization rate, cleavage rate and high-quality embryo rate from 21 cycles in the DC group (N=262 oocytes) and HC group (N=263 oocytes). Secondly, we determined the embryo transfer outcome and the offspring outcome in DC group (N=184 cycles) and HC group (N=136 cycles). Results: Compared with the HC group, significant increase was observed in the high-quality embryo rate (66.1.2% vs. 55.3%, p=0.037) and implantation rate (49.8% vs. 40.6%, p=0.027) in the DC group. No statistical differences were observed in the pregnant outcome and birth defect of the offspring (p>0.05). Compared with HC, DC was associated with a higher high-quality embryo rate and a higher implantation rate after embryo transfer. Conclusions: No statistical differences were noticed in the offspring conditions between the two culture modes. Taken together, DC may serve as a promising method for IVF-ET.

scholarly journals Preparation of cyclodextrin nanoparticles and evaluation of its effect on the capacitation of bovine spermatozoa used in the in vitro fertilization

2018 ◽  
Vol 9 (4) ◽  
pp. 112
Author(s):  
Heba F Salem ◽  
Mai Raslan ◽  
Hanaa Suliman ◽  
Tamer Essam ◽  
Saber Abd-Allah
Keyword(s):  
Data Analysis ◽  
In Vitro Fertilization ◽  
Interval Data ◽  
Fertilization Rate ◽  
Vitro Fertilization ◽  
Bovine Spermatozoa ◽  
The Media ◽  
Biomedical Interventions ◽  
Positive Effect

<p>This study was conducted to produce nanosized cyclodextrin (NCD) and assess its effect on bovine spermatozoa during In vitro fertilization (IVF) to optimize the capacitation media for successful IVF. Therefore, Four cyclodextrin formulations were prepared and characterized. Data analysis revealed the best formula (F2) showed a smallest particle size (15 nm), zeta potential (-37 mv), and higher yield percentages (95%) was selected for spem capacitation. Motile spermatozoa were separated from frozen-thawed semen by a swim-up procedure and capacitated in IVF-TALP medium with different formulae of NCD or CD or without treatments (control) and incubated for 3hours(hr) at 38°C and evaluated every one (hr) interval. Data analysis revealed that the formulation of cyclodextrin nanoparticles (F2<strong>)</strong> after (2hr) incubation in the media gave best effect on sperm capacitation and acrosme reaction (AR) and effect of sperm treated with NCD on fertilization rate was evaluated. The results showed that the proportion of Oocytes fertilized was increased significantly in F2 (60%) than in the control (35%), and cyclodextrin group (50%) groups (<em>p</em>&lt;0.05). It could be inferred from this investigation that cyclodextrin nanoparticles can be used for biomedical interventions in bovine spermatozoa. NCD improve sperm motility, viability, and (AR), also fertilization rate of sperm treated with NCD increase. So NCD gave positive effect on sperm functions during IVF. </p>

221 EFFECT OF ROCKING ON IN VITRO PRODUCTION OF BOVINE EMBRYOS

2009 ◽  
Vol 21 (1) ◽  
pp. 209
Author(s):  
Y. Serita ◽  
C. Kubota ◽  
T. Kojima
Keyword(s):  
In Vitro Fertilization ◽  
Embryo Development ◽  
Developmental Competence ◽  
In Vitro Production ◽  
Humid Atmosphere ◽  
Vitro Fertilization ◽  
Rate Of Development ◽  
Fetal Bovine ◽  
Vitro Production

This study tested whether embryo development yield using in vitro fertilization (IVF) could be improved by rocking cultures. Bovine ovaries were obtained at a slaughterhouse and transported to the laboratory within 6 h. Cumulus–oocyte complexes were collected and 20–25 were transferred in 100-μL drops of TCM-199 containing 10% fetal bovine serum and antibiotics under paraffin oil. Maturation was for 20–24 h at 38.5°C under 5% CO2 and 95% air in a humid atmosphere (IVM). In vitro fertilization was carried out for 6 h using frozen–thawed sperm from a single bull in modified Brackett and Oliphant (BO) medium. Presumptive zygotes were cultured in CR1aa supplemented with 10 mg mL–1 of BSA or 5% FBS for 9 d at 38.5°C under 5% CO2, 5% O2, and 90% N2 in a humid atmosphere (IVC). Rocking was performed to a height of 6 cm every 7 s using a Profile Rocker (New Brunswick Scientific Co., Edison, NJ, USA) in an incubator. Dishes were placed at a 15-cm distance from the fulcrum of the rocker. The conventional method (no rocking) served as a control, and every experiment was replicated 3 times. For Experiment 1, the effect of the period of rocking on developmental competence was examined when COC or zygotes were subjected to rocking for IVM, IVF, or IVC (IVM-move, IVF-move, and IVC-move). There were no significant differences in rates of oocyte maturation, cleavage, and development for IVM-move v. the control, or for rate of development between IVC-move and the control. However, the rate of fertilization for IVF-move was higher than that of the control (88.9 v. 67.5%; P < 0.01), and the rate of development was higher for IVF-move than for the control (39.0 v. 25.7%; P < 0.05). For Experiment 2, the effect of rocking frequency during IVF on development was determined. The IVF cultures were rocked every 7, 3.5, and 1.5 s (IVF-1move, IVF-2move, IVF-3move). The rates of cleavage on IVF-1move, IVF-2move, IVF-3move, and the control were 74.3, 69.8, 68.8, and 60.4%, and the rates of development were 39.0, 48.3, 26.2, and 25.7%, respectively. The rates of development on IVF-1move and IVF-2move were significantly different from the control and IVF-3move (P < 0.01). These results showed that rocking during IVF improved fertilization and embryo yield, and that frequency of rocking affected embryo development.

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