271 USE OF SEXED SEMEN FOR BOVINE IVF IN MEXICO: PRELIMINARY RESULTS

2006 ◽  
Vol 18 (2) ◽  
pp. 243
Author(s):  
J. Aysa ◽  
J. A. Medrano ◽  
A. Villa-Godoy ◽  
A. Barba ◽  
Y. C. Ducolomb ◽  
...  
Keyword(s):  
Sperm Concentration ◽  
Physiological Saline ◽  
Control Group ◽  
Concentration Gradients ◽  
Vitro Fertilization ◽  
Sexed Semen ◽  
Fort Collins ◽  
Vitro Development ◽  
Economic Improvement

The use of sexed embryos in dairy cattle is useful for the genetic and economic improvement of production. The aim of this study was to determine differences in in vitro maturation (IVM), in vitro fertilization (IVF), and in vitro development (IVD) with the use of sexed and unsexed sperm. Semen from one Holstein bull was used for the experiment. The semen was sexed and frozen by X-Y, Mexico (subsidiary of X-Y Inc., Fort Collins, CO, USA). Only the X fraction of spermatozoa was sorted and frozen in 0.25-mL plastic straws with 2.0 � 106 spermatozoa/straw. A modified protocol for IVF was used. A total of 334 ovaries were obtained from a local slaughterhouse and transported to the laboratory in physiological saline (25�C). From these, 1019 cumulus-oocyte complexes (COCs) were obtained and used for the procedures of IVM, IVF, and IVD. The average number of follicles aspirated per ovary was 3.6, and an average of 3.05 COCs were recovered per ovary. The oocyte recovery rate was 85%. For IVM, COCs were incubated in TCM-199 supplemented with BSA, pyruvate, FSH, and LH for 24 h. All incubations were performed at 38.5�C in a humidified atmosphere of 5% CO2 in air. After this period, COCs were placed in fertilization medium (TALP supplemented with BSA, heparin, penicillamine, and hypotaurine). For IVF, oocytes were randomly assigned to two groups: sexed semen (Holstein) or treatment group (TG), and non-sexed semen (Brahman, used as control in our laboratory) or control group (CG). For insemination, frozen-thawed semen from the Holstein and Brahman bulls was washed by centrifugation in two concentration gradients of a silicone solution. In both groups, sperm concentration used for IVF was 1 � 106 spermatozoa/mL. After insemination, oocytes and semen were co-incubated for 18 h. For IVD, presumptive zygotes were incubated for 7 days in a modified IVD medium (Barc-1) supplemented with BSA. The degree of IVD was evaluated according to the number of divisions of the embryos produced, and the number of embryos that developed to the morula and blastocyst stages. Five replicates were made. The rates of IVM, IVF, and IVD were analyzed by logistic regression. The COCs produced 890 fertilized oocytes. Of these, 442 were from the TG and 448 from the CG. A total of 393 embryos from the TG and 372 from the CG developed in vitro; embryos were evaluated for development on Day 7. A total of 108 morulae (21%) were produced in the TG and 100 (19%) in the CG, whereas 99 (19%) blastocysts developed in the TG and 105 (20%) in the CG. There were no statistically significant differences between the two groups studied for embryo IVD (P > 0.05). It is concluded that IVM, IVF, and IVD procedures used for conventional non-sexed semen can be used for similar results with sperm sexed by flow cytometry. This is the first report of sexed semen use for bovine IVF in Mexico and is a precedent for future investigations on in vitro embryo production in Mexico. More experiments are needed to confirm these preliminary findings. Sexed semen was provided by Rancho El Nacimiento, Establo 196, Tizayuca, State of Hidalgo, M�xico. Funding for J. A. was provided by CONACYT and UNAM.

scholarly journals 266IN VITRO-DERIVED EMBRYO PRODUCTION WITH SEXED AND UNSEXED SEMEN FROM DIFFERENT BULLS

2004 ◽  
Vol 16 (2) ◽  
pp. 253 ◽  
Author(s):  
L. Ferré ◽  
C. Ohlrichs ◽  
D. Faber
Keyword(s):  
In Vitro Fertilization ◽  
Embryo Development ◽  
Calf Serum ◽  
Sperm Concentration ◽  
Reproductive Technologies ◽  
Chi Square ◽  
Vitro Fertilization ◽  
Sexed Semen ◽  
Chi Square Analysis

The production of pre-sex-selected calves by in vitro fertilization (IVF), using sexed semen, does show some benefits due to the small quantity of sperms needed for the process as compared to other reproductive technologies. The objective of this study was to determine differences among bulls and sperm concentrations in embryo development with sexed and unsexed semen. Follicles ranging from 2 to 6mm in diameter were aspirated from slaughterhouse ovaries. COC were selected and matured in groups of maximum of 30 in 1.8mL of TCM-199, supplemented with 10% fetal calf serum, 0.01UmL−1 bFSH, 0.01UmL−1 bLH and 10μLmL−1 penicillin-streptomycin for 24h at 38.5°C. Fertilization (Day 0) was carried out in micro-drops (50μL) with TALP-FERT medium containing PHE (3μgmL−1 penicillamine, 11μgmL−1 hypotaurine and 0.18μgmL−1 epinephrine), 10μLmL−1 non-essential amino acid and 2μgmL−1 heparin. Frozen/thawed sexed (female) and non-sexed sperms from five bulls were selected in a discontinuous percoll gradient. Sperm concentration was 1×106 for non-sexed semen and 1×106 or 2×106 for sexed semen. After 18–20h, presumptive zygotes were denuded and cultured in groups of 10 in 50-μL micro-drops of SOF citrate with 5% FCS (Holm P et al., 1999 Theriogenology 52, 683–700) under paraffin oil in a 5% O2, 5% CO2, 90% N2 atmosphere with high humidity. On Day 7, blastocysts (BL) were morphologically evaluated and recorded. Results are shown in Table 1. Data was compared by chi-square analysis. Sexed frozen bovine sperm can be used successfully in IVF systems. More research needs to be done to optimize and standardize bovine in vitro fertilization with sexed semen. Table 1 Results of comparisons between bulls, sperm concentrations, cleavage and embryo development

171 IN VITRO FERTILIZATION USING FROZEN - THAWED SEXED SEMEN TREATED WITH RECOMBINANT HEPARIN-BINDING PROTEINS

2012 ◽  
Vol 24 (1) ◽  
pp. 197 ◽  
Author(s):  
E. A. Ordonez-Leon ◽  
M. E. Kjelland ◽  
J. F. Moreno ◽  
T. H. Welsh ◽  
R. D. Randel ◽  
...  
Keyword(s):  
Binding Proteins ◽  
Statistical Significance ◽  
Sperm Concentration ◽  
Tissue Inhibitor Of Metalloproteinase ◽  
Colloid Solution ◽  
System Response ◽  
Heparin Binding ◽  
Vitro Fertilization ◽  
Sexed Semen

The decrease in the fertility of males can have a negative economic impact on the production of milk and meat in cattle. As a result, interest exists in evaluating diverse proteins that might be able to increase the fertility of sperm. This is the case of the heparin-binding proteins (HBP), specifically the fertility-associated antigen (FAA) and the Type-2 tissue inhibitor of metalloproteinase (TIMP-2), which act to favour capacitation and the acrosome reaction, perhaps by modulating the immune system response toward the sperm. The objective of this study was to evaluate the effect of adding recombinant FAA and recombinant TIMP-2 to Hereford sexed semen (X-chromosome-bearing sperm) for use in an IVF program. For IVF, X-sorted semen was prepared both with and without HBP. For this, 25 μg of FAA and 25 μg of TIMP-2 were added per milliliter of semen and were left to adsorb for 20 min. The semen was then diluted, packaged in straws and cryopreserved. Ovaries were obtained from a slaughterhouse and transported to the laboratory to obtain cumulus–oocyte complexes by follicular aspiration and were cultured in maturation medium (TCM-199 supplemented with FSH, LH and oestradiol) for 24 h. All incubations were performed at 38.5°C in a humidified atmosphere of 5% CO2 in air. For insemination, frozen-thawed semen was washed by centrifugation in 2 concentration gradients of a silica-based colloid solution. The sperm concentration used for IVF was 1 × 106 spermatozoa mL–1. Gametes were co-incubated for 22 h in fertilization medium (TALP added with BSA, heparin, penicillamine, hypotaurine and epinephrine). For IVD, presumptive zygotes were incubated for 7 days in a modified IVD medium (SOF) supplemented with FCS and BSA. A total of 363 in vitro-matured oocytes were used in the control vs 405 oocytes in the treatment group. A Pearson chi-square test was used to determine the statistical significance. The cleavage rates for the control (61.4%) and treatment (83.7%) groups were different (P < 0.001). However, the numbers of blastocysts, expanded blastocysts and hatched blastocysts produced in the control (10.2, 6.3 and 0%, respectively) and treatment (7.4, 7.4 and 0.7%, respectively) groups were similar (all P > 0.1). The fact that there was an increase in the early embryonic development rates and not in the development of the embryos to the blastocyst stage suggests that HBP could play a role in fertilization, but not in embryo development. The results also demonstrate that the addition of the recombinant proteins to the semen increased the fertilizing capacity of the sperm at a concentration of 1 × 106 sperm cells mL–1 of fertilization medium. To further elucidate the effect of the HBP, an evaluation of sperm penetration and pronuclei formation is proposed as a complement to these results. We thank Tod C. McCauley, Roy L. Ax and the staff at TMI Laboratories International Inc. for their assistance. We also thank Richard W. Lenz and the staff at Sexing Technologies for their assistance.

scholarly journals The effects of supplemented sericin on in vitro maturation and preimplantation development of mouse embryos: An experimental study

2021 ◽  
pp. 921-928
Author(s):  
Omid Banafshi ◽  
Sherko Nasseri ◽  
Leila Farhadi ◽  
Masoud Alasvand ◽  
Mohammad Bagher Khadem-Erfan ◽  
...  
Keyword(s):  
Embryo Culture ◽  
In Vitro Maturation ◽  
Culture Medium ◽  
Control Group ◽  
Vitro Fertilization ◽  
In Vitro Development ◽  
Embryo Culture Medium ◽  
Vitro Development

Background: Mouse embryo culture condition is an essential part of transgenic, reproductive and developmental biology laboratories. Mouse embryonic culture media may have a high risk of serum contamination with pathogens.  Objective: To investigate the effect of sericin as an embryo culture medium supplement on in vitro maturation (IVM), in vitro fertilization (IVF), and development of the preimplantation embryo in mice. Materials and Methods: The effects of sericin at three concentrations (subgroups) of 0.1%, 0.5%, and 1% as a medium supplement on IVM, IVF, and in vitro development of mouse embryos were separately investigated and compared with a sericin-free (control) group. The cumulative effect of the three concentrations was evaluated for IVM + in vitro development and IVF + in vitro development as follow-up groups. Results: In the IVM group, compared to the control group, the number of oocysts reaching the MII stage was significantly higher when 1% sericin was used (161/208 = 77.4%). No significant results were observed in the IVF and in vitro development groups with different concentrations of sericin compared to the control group. Among the follow-up groups, in the IVM + in vitro development group, the number of oocytes was higher after passing the IVM and IVF and reaching the blastocysts stage when 1% sericin was used, compared with other sericin subgroups. A significant difference was also noted when compared with the control group (p = 0.048). The IVF + in vitro development study group, on the other hand, did not show any significant relationship. Conclusion: It can be concluded that 1% sericin can be used as a supplement in mouse embryo cultures to improve the IVM rate. Also, based on the findings, sericin appears to be an effective supplement which can have a positive effect on the development of embryos derived from IVM. Key words: Sericin, In vitro maturation, In vitro fertilization, Preimplantation embryo, Culture medium, Mice.

Levels interleukine-4 and interleukin-17 (IL-4, IL-17) of blood in patients with habitual recurrent pregnancy loss, which occurred in a loop in vitro fertilization

2016 ◽  
pp. 137-139
Author(s):  
K.P. Golovatyuk ◽  
Keyword(s):  
In Vitro Fertilization ◽  
Conditioned Medium ◽  
Mononuclear Cells ◽  
Recurrent Miscarriage ◽  
Interleukin 4 ◽  
Control Group ◽  
Interleukin 17 ◽  
Vitro Fertilization ◽  
Blood Mononuclear Cells

The objective: was to investigate the levels of cytokines IL-4 and IL-17 in serum and conditioned medium cultures of blood mononuclear cells (MNC) and evaluation association between their products and miscarriage, which occurred in IVF cycles. Patients and methods. We observed 240 patients with recurrent miscarriage, came in IVF cycles, and 100 apparently healthy fertile women in the control group. The concentrations of IL-4 and IL-17 in serum and conditioned medium of MNC cultures were determined. Results. The levels of IL-4 in the serum and conditioned medium in spontaneous and stimulated mitogen secretion was not significantly different from those in the control group, whereas IL-17 levels were higher than those in the control group serum, in conditioned media of stimulated and non-stimulated MNCs. Conclusion. Disregulation of activity of circulating blood mononuclear cells in women with recurrent miscarriage that followed IVF, is accompanied by increased secretion of IL-17 and almost constant production of IL-4 on the back of high stimulation index of production of these cytokines. Key words: in vitro fertilization, miscarriage, interleukin-4, interleukin-17, serum stimulated and non-stimulated mononuclear blood.

scholarly journals 252 COMPARISON OF TWO SPERM SEPARATION PRODUCTS FOR USE IN BOVINE IVF

2005 ◽  
Vol 17 (2) ◽  
pp. 276 ◽  
Author(s):  
J. Pryor ◽  
S. Romo ◽  
D.D. Varner ◽  
K. Hinrichs ◽  
C.R. Looney
Keyword(s):  
Sperm Concentration ◽  
Embryo Production ◽  
Chi Square ◽  
Vitro Fertilization ◽  
Before And After ◽  
Chi Square Test ◽  
Group 2 ◽  
Live Sperm ◽  
Sperm Separation

In commercial bovine in vitro fertilization (IVF) companies, there is a continuous need to improve results. Efforts to maximize in vitro embryo production have included modifications in the use of sperm separation gradients. The development of commercially available sperm centrifugation gradients represents a new possibility of increasing the number of viable sperm that can be obtained from low concentration (fresh or frozen, sexed or unsexed) semen samples in order to improve the efficiency of the IVF system to make embryo production as efficient as possible. The objective of this study was to compare two different separation gradients, as follows: Group 1: Percoll (Sigma, St. Louis, MO, USA), in 45% and 90% gradients; Group 2: EquiPure (Nidacon, Gathenburg, Sweden), in top and bottom layers. Before and after separation, sperm were evaluated at 200× magnification for total motility, and then stained to assess viability at 400× with fast-green/eosin stain (Sigma). Sperm separation was performed using frozen/thawed semen from one bull. Semen was separated by centrifugation at 200g for 30 min in both density gradients. Results obtained from Groups 1 and 2 were compared by chi-square test. Sperm separation with Percoll yielded lower numbers of sperm (average sperm concentration after separation of 92 × 106, vs. 159 × 106 sperm/mL for EquiPure; P < 0.05) but resulted in higher motility (60% vs. 39%, respectively; P < 0.05) of separated sperm. Rates of live sperm cells were not significantly different between groups (69.5% vs. 70%, respectively; P > 0.1). These results indicate that the commercial separation medium EquiPure may be associated with higher sperm concentration levels but with lowered sperm motility when compared to Percoll for bovine sperm separation. However, Equipure provided similar percentages of live sperm when compared to Percoll, which is currently used in our laboratory.

Calibration of sperm concentration for in vitro fertilization in a mouse reprotoxicity model

2019 ◽  
Vol 55 ◽  
pp. 58-61 ◽  
Author(s):  
Hanne Skovsgaard Pedersen ◽  
Ying Liu ◽  
Leslie Foldager ◽  
Henrik Callesen ◽  
Knud Larsen ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Sperm Concentration ◽  
Vitro Fertilization

scholarly journals IN VITRO FERTILITY TEST OF HUMAN SPERMATOZOA MEMBRANE PROTEIN FERTILIN BETA ANTIBODY IN MICE (Mus musculus Balb/c) AS IMMUNOCONTRACEPTIVE CANDIDATE

2017 ◽  
Vol 52 (3) ◽  
pp. 209
Author(s):  
Reny I’tishom ◽  
Doddy M Soebadi ◽  
Aucky Hinting ◽  
Hamdani Lunardhi ◽  
Rina Yudiwati
Keyword(s):  
In Vitro Fertilization ◽  
Membrane Protein ◽  
Mus Musculus ◽  
Human Spermatozoa ◽  
Control Group ◽  
Reproductive Process ◽  
Vitro Fertilization ◽  
Antibody Induction

One of the materials as potential candidates immunocontraception material is spermatozoa. Fertilin beta is spermatozoa membrane protein and is found only in mature spermatozoa and ejaculate, which serves as an adhesion molecule. Spermatozoa membrane protein that is used as an ingredient immunocontraception candidate, must have specific criteria that the specificity of spermatozoa, the role of antigen in the fertilization process, which includes the formation of immunogenicity sufficient antibody response has the potential to block fertilization. Antibodies against spermatozoa affect the stages before fertilization of the reproductive process and can hinder the development of the embryo after fertilization. Until now very little research data spermatozoa membrane protein as an ingredient immunocontraception are up to the test of experimental animals. The research objective is to prove the role of the resulting antibody induction of antibodies fertilin beta protein in the membrane of human spermatozoa induce agglutination and reduce motility thus reducing the number of in vitro fertilization. Research conducted at the IVF Laboratory, Department of Biology of Medicine, Faculty of Medicine, University of Airlangga. This research includes: Test the potential of antibody protein beta fertilin membrane of human spermatozoa and inhibit the role of antibodies in vitro fertilization in mice (Mus musculus Balb/c). In vitro studies have resulted in fertilization figure of 25% is smaller than the number that is equal to control fertilization of 58.7%, whereas previously the spermatozoa were incubated first with a beta membrane protein antibody fertilin human spermatozoa. While the percentage of inhibition of sperm to fertilize an oocyte by 33.75%. Potential imunokontraseptif considered effective if it decreased significantly (P <0.05) than the numbers fertilization in the treatment group compared with the control group. This shows fertilin beta membrane protein antibody has the ability to inhibit human spermatozoa to fertilize oocytes that reduce the number of fertilization.

scholarly journals The effectiveness of collaborative infertility counseling (CIC) on pregnancy outcome in infertile women undergoing in vitro fertilization: A randomized controlled trial

2019 ◽  
Author(s):  
Mahboobeh Rasoulzadeh Bidgoli ◽  
robab latifnejad roudsari ◽  
ali montazeri
Keyword(s):  
In Vitro Fertilization ◽  
Pregnancy Rate ◽  
Treatment Success ◽  
Intervention Group ◽  
Control Group ◽  
Vitro Fertilization ◽  
Infertile Women ◽  
Significant Difference ◽  
And Control

Abstract Background: Infertility is an emotional tension which influences the whole aspects of relationships in infertile couples. A main objective of infertility treatments is elevation of pregnancy rate. The present study aimed to examine the effect of collaborative counseling on pregnancy rate in infertile women, undergoing in vitro fertilization in Mashhad, Iran. Methods: In this clinical trial, 60 women with primary infertility were selected from an infertility research center and were randomly allocated into intervention (n=29) and control (n=31) groups. The intervention group received individual counseling, based on the collaborative reproductive healthcare model with collaboration of a midwife, a gynecologist and a clinical psychologist in five sessions during a two-month period. The control group received routine care. Positive pregnancy test was considered as a criterion of treatment success at the end of the study. Data were analyzed using statistical tests including independent samples t-test. Results: There was no significant difference in pregnancy rate between intervention and control groups (P = 0.298). Also, there were no significant differences in follicle and embryo numbers between two groups. However, a significant difference was observed between two groups in terms of oocyte numbers where the intervention group had more oocyte (P = 0.014). Conclusion: Overall the findings indicated that the collaborative infertility counseling did not improve treatment success in infertile women undergoing in vitro fertilization

scholarly journals Piglets produced by transfer of embryos obtained by in vitro fertilization of oocytes matured in vitro with thymosin: A case report

2020 ◽  
Vol 76 (03) ◽  
pp. 6356-2020 ◽  
Author(s):  
KATARZYNA PONIEDZIAŁEK-KEMPNY ◽  
BARBARA GAJDA ◽  
IWONA RAJSKA ◽  
LECHOSŁAW GAJDA ◽  
ZDZISŁAW SMORĄG
Keyword(s):  
Case Report ◽  
In Vitro Fertilization ◽  
Control Group ◽  
First Birth ◽  
Vitro Fertilization ◽  
Research Material ◽  
Preliminary Study ◽  
Experimental Group

The aim of the study was to examine the in vivo viability of in vitro-produced (IVP) porcine embryos obtained from oocytes matured with thymosin. The research material for this study consisted of immature pig oocytes obtained from ovaries after slaughter and ejaculated semen obtained from one boar. The immature oocytes were cultured in vitro until the metaphase II stage in a medium supplemented with thymosin (TMS). The presumptive zygotes obtained were cultured in vitro for 4-40 hours. The presumptive zygotes and 2-4-cell embryos were evaluated in vivo after transferring them to synchronized recipients. After the transfer of embryos from the experimental group into 2 recipients (50 embryos into each gilt) and the transfer of 50 embryos from the control group into 1 recipient, both gilts that had received embryos obtained by in vitro fertilization of oocytes matured with TMS became pregnant and delivered a total of 16 live piglets. After the transfer of embryos from the control group, no pregnancy was achieved. In conclusion, the results of our preliminary study suggest that the maturation of pig oocytes with thymosin supports the in vivo survival of in vitro produced embryos. It is important to note, that this was the first birth of piglets obtained after transfer of IVP embryos in Poland.

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