Comparison between the in vitro fertilization capability of frozen semen from different bulls

1999 ◽  
Vol 51 (1) ◽  
pp. 328
Author(s):  
G Rosés ◽  
C Larocca ◽  
I Lago ◽  
J Calvo ◽  
M Viqueira ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Vitro Fertilization ◽  
Frozen Semen

PSXI-10 How to use ultrasonic cutter on frozen semen for multiple embryo production

2021 ◽  
Vol 99 (Supplement_3) ◽  
pp. 248-249
Author(s):  
Sung Woo Kim ◽  
Jae-Yeong Lee ◽  
Chan-Lan Kim ◽  
In-Sul Hwang ◽  
Yeoung-Gyu Ko ◽  
...  
Keyword(s):  
Survival Rate ◽  
In Vitro Fertilization ◽  
Endangered Species ◽  
Genetic Resources ◽  
Bovine Embryos ◽  
Vitro Fertilization ◽  
Economical Method ◽  
Frozen Semen ◽  
Ultrasonic Cutting

Abstract The objective of the present study was to establish conditions for using technology that can potentially enhance the efficiency of bovine embryos derived from in vitro fertilization (IVF) with frozen semen. Frozen semen from selected bulls can be stored indefinitely in liquid nitrogen as genetic resources; however, these resources are considered consumable because they cannot be regenerated. Therefore, to optimize the utilization of frozen semen, as many oocytes as possible should be fertilized with one straw. However, a sufficient number of prepared oocytes might not be available for one experiment, which can limit the use of the total spermatozoa population. Thus, an economical method for producing embryos needs to be established by optimizing technology for transplantable embryos. In this study, the utilization of frozen semen was increased by dividing the straw with an ultrasonic cutter. The post-thaw survival rate of uncut straws from Korean Proven Bulls did not differ from that of half cuttings. When ultrasonic cutting was applied to frozen semen, spermatozoa could be prepared for IVF trials at least four times, and blastocysts were produced. Therefore, cutting frozen semen with an ultrasonic cutter represents a potentially useful tool to expand genetic resources from excellent breeding stocks. This approach could also be valuable in the field of IVF of endangered species or rare breeds for their preservation, as well as in ovum pick-up (OPU) techniques.

Is fresh or frozen semen to be used in in vitro fertilization with donor sperm?

1989 ◽  
Vol 51 (4) ◽  
pp. 661-664 ◽  
Author(s):  
Yvon. Englert ◽  
Annick. Delvigne ◽  
Marcel. Vekemans ◽  
Bernard. Lejeune ◽  
Alain. Henlisz ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Donor Sperm ◽  
Vitro Fertilization ◽  
Frozen Semen

Role of back-up frozen semen in couples undergoing in vitro fertilization (IVF)

2004 ◽  
Vol 82 ◽  
pp. S265
Author(s):  
A.B. Pinto ◽  
M.J. Putman ◽  
S. Marynick ◽  
N. Broyles ◽  
R.D. Dickerson ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Vitro Fertilization ◽  
Frozen Semen

365 COMPARISON OF IN VITRO FERTILIZING CAPACITY OF FROZEN - THAWED SEX-SORTED AND SEX-SORTED FROZEN - THAWED BULL SPERMATOZOA

2007 ◽  
Vol 19 (1) ◽  
pp. 298 ◽  
Author(s):  
V. Malcolm ◽  
M. Marfil ◽  
M. Calvi ◽  
F. Rigali ◽  
M. Pugliese ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Embryo Development ◽  
Egg Yolk ◽  
Reproductive Technologies ◽  
Development Rate ◽  
Vitro Fertilization ◽  
Final Treatment ◽  
Frozen Semen ◽  
Sexed Semen

Sperm sexing has become a world-wide technology, now available in many countries. The method has been incorporated into many reproductive technologies such as embryo production (Zhang et al. 2003 Theriogenology 60, 1657–1663), but sex-sorting is limited when bulls are located far from sorters or when only frozen semen is available. Previous studies on sexing frozen–thawed spermatozoa have been done in rams, which resulted in retention of the spermatozoan functional capacities (Hollinshead et al. 2004 Reproduction 127, 557–568). In vitro characteristics were assessed in bulls after sexing of thawed sperm (Hollinshead et al. 2004 Theriogenology 62, 958–968); however, the fertilizing capacity of frozen–thawed sex-sorted (FTSS) spermatozoa was not tested. The aim of the present study was to compare cleavage and embryo development rate among frozen–thawed (FT), sex-sorted frozen–thawed (SSFT), and FTSS bull spermatozoa. For FT, sperm were diluted to a final concentration of 60 × 106 sperm/mL, packaged in 0.5-mL straws, and frozen. In SSFT, spermatozoa were sex-sorted by flow cytometry following Schenk protocols (1999 Theriogenology 52, 1375–1391). Three × 106 spermatozoa were packaged into 0.25-mL straws and frozen. The final treatment (FTSS) consisted of thawing 6 to 10 frozen straws of 4 different bulls containing an average of 25 × 106 spermatozoa and centrifuging at 600g for 15 min at 21°C to extract cryodiluent. Spermatozoa were diluted and stained with Hoechst 33342 (stain concentration of 112.5 µM, the same used for SSFT treatment) following Schenk sexed-semen protocols (1999), sex-sorted by a flow cytometer, and collected in Tris-base extender containing 20% egg yolk. For each ejaculate frozen–thawed, SSFT and FTSS spermatozoa were prepared for oocyte in vitro fertilization. Also, semen from a bull routinely used as a control in the laboratory was added for a better comparison of results. Oocytes from a slaughterhouse were processed following standard in vitro fertilization procedures (Ferré 2002 Theriogenology 57, 664) 4 times for each bull, and comparison was made between treatments. Results were analyzed by ANOVA. No significant differences were observed among bulls (data not shown) (P > 0.05). Although embryo development rate was statistically different between sexed and non-sexed groups (P < 0.05), results showed that frozen–thawed bull spermatozoa can be sex-sorted and used for in vitro fertilization with comparable developmental rates comparable to those when frozen sexed semen is used (Table 1). This opens a new commercial window for cases where pre-selected sexed embryos from bulls that are not in AI centers are desired, also giving an opportunity for dead bulls. Nevertheless, since a large number of straws are necessary, further studies must be carried out to make this procedure more efficient and economically profitable. Table 1. Results of cleavage and embryo development rates between spermatozoa treatments

181 IN VITRO FERTILIZATION (IVF) USING SEMI-DEFINED CULTURE CONDITIONS FROM LOW OR HIGH ANTRAL FOLLICLE COUNT PUBERTAL BEEF HEIFERS

2016 ◽  
Vol 28 (2) ◽  
pp. 221
Author(s):  
C. C. Chase ◽  
R. A. Cushman ◽  
A. K. McNeel ◽  
O. L. Amundson ◽  
G. A. Perry ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Prostaglandin F2α ◽  
Antral Follicle ◽  
Antral Follicle Count ◽  
Oestrous Cycle ◽  
Vitro Fertilization ◽  
Development Rates ◽  
Frozen Semen ◽  
Defined Culture

To compare the in vitro fertilization (IVF) and production (IVP) of embryos from low and high antral follicle count (AFC) heifers, AFC were determined on 106 heifers using transrectal ultrasonography. Ten heifers with the lowest AFC (avg. 13.2) and 10 heifers with the highest AFC (avg. 27.4) with evidence of oestrous cyclicity were synchronised with 2 injections of prostaglandin F2α (PGF2α); half were harvested on Days 5 to 6 and half on Days 15 to 16 of the oestrous cycle. The IVF procedures included protocols for semi-defined media and were as described (IVP Protocol, P. J. Hansen’s Laboratory, University of Florida, Gainesville, FL, USA). Cumulus-oocyte complexes (COC) from follicles less than 8 mm in diameter were cultured in maturation medium (5% CO2; 38.5°C) for 24 h. Matured COC were fertilised using thawed frozen semen from a crossbred bull that was purified using Percoll gradient separation procedures. Motile spermatozoa were added to COC in fertilization medium at a final concentration of 1 × 106 spermatozoa per mL. About 24 h later, presumptive zygotes were placed in microdrops of development medium under oil, and cultured (5% CO2, 5% O2, balance N2, 38.5°C). On Days 3 and 8 after fertilization, cleavage and blastocyst development rates, respectively, were assessed. Data were analysed using the mixed procedure of SAS (SAS Institute, Cary, NC, USA) and the model included the effects of collection day, group (high or low AFC), and their interaction. More COC (P < 0.0005) were collected from high than low AFC heifers (30.3 v. 9.3 ± 3.12 per heifer). Both the number and percentage of COC that cleaved had an interaction between collection day and group (P < 0.03). The interaction appeared to be due to low cleavage and development rates on 1 of 4 collection days (appeared not related to day of oestrous cycle). Although high compared to low AFC heifers had more COC that cleaved (18.7 v. 4.4 ± 1.84 per heifer), the percentage of cleaved COC did not differ (59.2 v. 49.8 ± 3.36%). There were no significant differences between high and low AFC heifers in the number of blastocysts (3.1 v. 0.6 ± 1.21 per heifer) or in the percentage of COC that developed to blastocysts (8.8 v. 5.2 ± 3.60%). In previous replicates (years), we reported that high AFC heifers had more COC collected, more COC that cleaved, and more COC that developed to blastocysts than low AFC heifers. In contrast, in this study numbers of COC that developed to blastocysts did not significantly differ between high and low AFC heifers. Additionally, the percentage of COC that cleaved, and that developed to blastocyst have been similar between high and low AFC heifers. Therefore, high compared to low AFC heifers may produce more IVP embryos; however, AFC does not appear to affect the competence of an oocyte to develop and mature to the blastocyst stage.

scholarly journals Ultrasonic Cutting of Frozen Semen Straws to Optimize the Use of Spermatozoa for In Vitro Fertilization

Animals ◽  
2020 ◽  
Vol 10 (11) ◽  
pp. 2152
Author(s):  
Sung Woo Kim ◽  
Jae-Yeong Lee ◽  
Bongki Kim ◽  
Chan-Lan Kim ◽  
In-Sul Hwang ◽  
...  
Keyword(s):  
Survival Rate ◽  
In Vitro Fertilization ◽  
Endangered Species ◽  
Genetic Resources ◽  
Bovine Embryos ◽  
Vitro Fertilization ◽  
Economical Method ◽  
Frozen Semen ◽  
Ultrasonic Cutting

The objective of the present study was to establish conditions for using technology that can potentially enhance the efficiency of bovine embryos derived from in vitro fertilization (IVF) with frozen semen. Frozen semen from selected bulls can be stored indefinitely in liquid nitrogen as genetic resources; however, these resources are considered consumable because they cannot be regenerated. Therefore, to optimize the utilization of frozen semen, as many oocytes as possible should be fertilized with one straw. However, a sufficient number of prepared oocytes might not be available for one experiment, which can limit the use of the total spermatozoa population. Thus, an economical method for producing embryos needs to be established by optimizing technology for transplantable embryos. In this study, the utilization of frozen semen was increased by dividing the straw with an ultrasonic cutter. The post-thaw survival rate of uncut straws from Korean Proven Bulls did not differ from that of half cuttings. When ultrasonic cutting was applied to frozen semen, spermatozoa could be prepared for IVF trials at least four times, and blastocysts were produced. Therefore, cutting frozen semen with an ultrasonic cutter represents a potentially useful tool to expand genetic resources from excellent breeding stocks. This approach could also be valuable in the field of IVF of endangered species or rare breeds for their preservation, as well as in ovum pick-up (OPU) techniques.

scholarly journals 249 EFFECT OF REFREEZING BULL SEMEN ON IVF SUCCESS RATE

2005 ◽  
Vol 17 (2) ◽  
pp. 275
Author(s):  
P. McCue ◽  
J. Kelly ◽  
S. Ashworth ◽  
D. Kleemann ◽  
S. Walker
Keyword(s):  
In Vitro Fertilization ◽  
Assisted Reproduction ◽  
Blastocyst Development ◽  
Vitro Fertilization ◽  
Colorado State University ◽  
Treatment Groups ◽  
Bull Semen ◽  
Frozen Semen ◽  
Group 2

Cryopreserved semen from valuable sires may be available in limited quantities in some situations. A large percentage of the spermatozoa in a thawed straw is potentially wasted since a relatively small number of spermatozoa are required for most assisted reproduction techniques. The goal of this study was to determine the effect of dilution and refreezing of bull semen on fertilization and blastocyst development rates following in vitro fertilization. The hypothesis was that frozen bull semen that was thawed, diluted, and refrozen could be used successfully for IVF. Oocytes were harvested from cow ovaries collected from a local abattoir and matured in vitro for 24 hours. Ova were subsequently assigned to one of four in vitro fertilization treatment groups. Group 1 ova (n = 158) were fertilized with bull semen frozen at a concentration of 20 × 106 spermatozoa per 0.25 mL straw. Group 2 ova (n = 157) were fertilized with semen frozen at an initial concentration of 2 × 106 spermatozoa. Group 3 ova (n = 157) were fertilized with semen that had been thawed and refrozen at a concentration of 20 × 106 spermatozoa. Group 4 ova (n = 150) were fertilized with semen that had initially been frozen at a concentration of 20 × 106 spermatozoa and then thawed, diluted to a concentration of 2 × 106 spermatozoa, and refrozen. IVF was performed in a medium volume of 100 μL using 1 × 106 spermatozoa/mL. Cleavage and blastocyst rates were determined 2 days and 7 days, respectively after IVF. Cleavage rates following IVF was highest with semen frozen at 20 × 106 spermatozoa (89.9%), intermediate with semen frozen at 2 × 106 spermatozoa or refrozen at 20 × 106 spermatozoa (71.3% and 73.9%, respectively), and lowest with semen refrozen at 2 × 106 spermatozoa (38.7%) (P < 0.05). Blastocyst development rate was not significantly different (P > 0.05) among treatment groups. This study confirmed the hypothesis that refrozen bovine semen can be used successfully for in vitro fertilization. Although the overall IVF efficiency was lower using diluted refrozen semen, multiple IVF procedures could theoretically be performed over time from one initial straw. Consequently, if a limited amount of frozen semen is available, thawing of a single straw followed by dilution, re-allocation into multiple straws, and refreezing should be considered to facilitate the more efficient use of semen in future assisted reproduction endeavors. This study was supported by the PEG Program, Colorado State University.

222 THE USE OF IN VITRO FERTILIZATION TO STUDY BOVINE IDIOPATHIC INFERTILITY

2009 ◽  
Vol 21 (1) ◽  
pp. 209
Author(s):  
L. G. B. Siqueira ◽  
C. W. Palmer ◽  
C. Lessard
Keyword(s):  
In Vitro Fertilization ◽  
Sperm Motility ◽  
Negative Control ◽  
Blue Staining ◽  
Computer Assisted ◽  
Vitro Fertilization ◽  
Coomassie Blue ◽  
Frozen Semen ◽  
Fresh Semen

A 4-year-old beef bull underwent a breeding soundness evaluation at the Western College of Veterinary Medicine (Veterinary Hospital, University of Saskatchewan); no apparent abnormalities were observed after conventional semen evaluations. However, the clinical history of this bull indicated that no pregnancies resulted from natural service of 52 cycling cows over a period of 2 years. Completed services were observed. The objective of this study was to use in vitro fertilization (IVF) technology to evaluate whether the sperm from this infertile bull could successfully fertilize oocytes in vitro. Fresh semen was collected with an electroejaculator and frozen in a computer-controlled rate freezer. Concentration and motility parameters were assessed by using computer-assisted semen analyses. Sperm morphology was evaluated on eosin-nigrosin-stained slides, and Coomassie blue staining was used to evaluate the presence of intact acrosomes. Within each evaluation technique, frozen–thawed semen from bulls (n = 2) with proven fertility was used as a positive control and samples of dead sperm (produced by repeated frozen–thawed cycles until reaching no sperm motility) were used as a negative control. Frozen semen from the infertile bull was used for IVF assay. Data were analyzed by ANOVA (sperm motility and the presence of acrosomes) or the chi-square test (cleavage and blastocyst rates), with a P-value of 0.05. Our infertile bull showed an average motile sperm percentage of 91.7 ± 2.2%, with 78.6 ± 3.5% progressive motility. After cryopreservation procedures, frozen–thawed semen had an acceptable general and progressive percentage of motility of 57.8 ± 6.7% and 43.9 ± 9.2%, respectively. Sperm stained with Coomassie blue showed a greater proportion of intact acrosomes in the fresh semen (63.6 ± 4.3% v. 40.4 ± 3.7%; P < 0.05); however, frozen–thawed semen from both the fertile bull and the control were similar (40.4 ± 3.7b% v. 45.5 ± 2.2b%, P < 0.05). In vitro fertilization results revealed a low cleavage rate at 48 h postfertilization (19.8 ± 6.3%) and blastocyst rate (2.4 ± 2.8%) when using frozen–thawed semen from the infertile bull, compared with the control (56.7 ± 8.2% and 26.3 ± 4.5%, respectively; P < 0.001). Moreover, cleavage and blastocyst rates obtained by using the negative control (21.1 ± 3.2% and 1.1 ± 1.9%, respectively) were similar to those of the infertile bull (P > 0.10). It was noted that ova fertilized with either frozen–thawed semen from the infertile bull or the negative control did not produce blastocysts before Days 8 and 9 of embryo culture, which is a characteristic of parthenogenesis. The results from this IVF study suggest that in this bull, there was a missing or defective factor blocking one of the steps in the fertilization process. Further investigations to identify this factor will increase our knowledge of male fertility, and could lead to new methods of evaluating and regulating fertilizing ability.

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