scholarly journals Collection, freezing of Ban epididymal sperm and evaluation of in vitro fertility after thawing

2021 ◽  
Vol 18 (4) ◽  
pp. 625-632
Author(s):  
Nguyen Viet Linh ◽  
Nguyen Thi Nhung
Keyword(s):  
In Vitro Fertilization ◽  
Sperm Concentration ◽  
Epididymal Sperm ◽  
Freezing And Thawing ◽  
Vitro Fertilization ◽  
Mountainous Regions ◽  
Before And After ◽  
Thawing Processes ◽  
Motility Of Sperm

Many researches on characteristics of porcine ejaculated semen on popular commercialized breeds collecting at breeding centers or households with pig breeding services have been carried out in Vietnam. However, very few investigation had been dealt with epididymal sperms, especially the ones from boar of Ban, a popular Vietnam native mini-pig breed which is usually freely raised in far rural and mountainous regions in some provinces of Northern Vietnam. In the present study, we surveyed on criteria of epididymal sperms from 4 Ban boars at collection, after freezing and thawing processes, and tested their fertility by an in vitro fertilization and embryo culture experiment. Approximate volume of sperm collected from a Ban boar did not differ (3-4 mL), with sperm concentration from 6.4 x 109 to 11.3 x 109 sperm/mL. Motility varied from 8.7 to 27.0%, whereas vitality was from 58.0 to 85.6%. After freezing and thawing, the motility of sperm slightly decreased to values of 6.3 to 25.7%, and viability significantly decreased to values of 41.3 to 79.6%. No difference was found between rates of abnormal morphology before and after freezing and thawing (10.6 to 31.0% and 12.0 to 32.0%, respectively). A test by in vitro fertilization with Landrace oocytes revealed that two sperm lots had acceptable in vitro fertility with rates of blastocyst formation from 14.4 to 18.8%. In conclusion, a study on collection, freezing and fertility testing of epididymal sperm collected from Ban boars has been carried out. The results of the present study could contribute necessary information as well as standardized sperm lots as important materials to further research on in vitro fertilization of Ban in Vietnam.

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 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

scholarly journals Incidence of Y-chromosome microdeletions in children whose fathers underwent vasectomy reversal or in vitro fertilization with epididymal sperm aspiration: a case-control study

2016 ◽  
Vol 14 (4) ◽  
pp. 534-540 ◽  
Author(s):  
Milton Ghirelli-Filho ◽  
◽  
Patricia Leme de Marchi ◽  
Fernanda Abani Mafra ◽  
Viviane Cavalcanti ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Y Chromosome ◽  
Epididymal Sperm ◽  
Sperm Retrieval ◽  
Vitro Fertilization ◽  
Y Chromosome Microdeletions ◽  
Sperm Aspiration ◽  
Vasectomy Reversal ◽  
Control Study

ABSTRACT Objective To evaluate the incidence of Y-chromosome microdeletions in individuals born from vasectomized fathers who underwent vasectomy reversal or in vitro fertilization with sperm retrieval by epididymal aspiration (percutaneous epididymal sperm aspiration). Methods A case-control study comprising male children of couples in which the man had been previously vasectomized and chose vasectomy reversal (n=31) or in vitro fertilization with sperm retrieval by percutaneous epididymal sperm aspiration (n=30) to conceive new children, and a Control Group of male children of fertile men who had programmed vasectomies (n=60). Y-chromosome microdeletions research was performed by polymerase chain reaction on fathers and children, evaluating 20 regions of the chromosome. Results The results showed no Y-chromosome microdeletions in any of the studied subjects. The incidence of Y-chromosome microdeletions in individuals born from vasectomized fathers who underwent vasectomy reversal or in vitro fertilization with spermatozoa recovered by percutaneous epididymal sperm aspiration did not differ between the groups, and there was no difference between control subjects born from natural pregnancies or population incidence in fertile men. Conclusion We found no association considering microdeletions in the azoospermia factor region of the Y chromosome and assisted reproduction. We also found no correlation between these Y-chromosome microdeletions and vasectomies, which suggests that the assisted reproduction techniques do not increase the incidence of Y-chromosome microdeletions.

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