scholarly journals Comparison of Viability, ATP and In vitro Fertilization of Boar Sperm Stored at 4??C in the Three Different Diluents

2008 ◽  
Vol 21 (8) ◽  
pp. 1127-1133
Author(s):  
Y. J. Yi ◽  
Z. H. Li ◽  
E. S. Kim ◽  
E. S. Song ◽  
P. Q. Cong ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Vitro Fertilization ◽  
Boar Sperm

Effects of Isatis root polysaccharide on boar sperm quality during liquid storage and in vitro fertilization

2019 ◽  
Vol 210 ◽  
pp. 106178 ◽  
Author(s):  
Zhiqiang Ren ◽  
Weike Shaoyong ◽  
Qian Li ◽  
Lu Ma ◽  
Junying Xiao ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Sperm Quality ◽  
Vitro Fertilization ◽  
Liquid Storage ◽  
Boar Sperm

Effect of Astragalus polysaccharide addition to thawed boar sperm on in vitro fertilization and embryo development

2018 ◽  
Vol 121 ◽  
pp. 21-26 ◽  
Author(s):  
Xiao-gang Weng ◽  
Ming-ming Cai ◽  
Yu-ting Zhang ◽  
Yan Liu ◽  
Zheng-ling Gao ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Embryo Development ◽  
Vitro Fertilization ◽  
Astragalus Polysaccharide ◽  
Boar Sperm

Factors affecting homologous in vitro fertilization assay of boar sperm fertility

1994 ◽  
Vol 41 (1) ◽  
pp. 249
Author(s):  
C. Matás ◽  
E. Martínez ◽  
J.M. Vázquez ◽  
J. Roca ◽  
J. Gadea ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Vitro Fertilization ◽  
Factors Affecting ◽  
Boar Sperm

scholarly journals Liquid Boar Sperm Quality during Storage and In vitro Fertilization and Culture of Pig Oocytes

2004 ◽  
Vol 17 (10) ◽  
pp. 1369-1373 ◽  
Author(s):  
C. S. Park ◽  
M. Y. Kim ◽  
Y. J. Yi ◽  
Y. J. Chang ◽  
S. H. Lee ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Sperm Quality ◽  
Vitro Fertilization ◽  
Boar Sperm

scholarly journals 289EFFECT OF FERTILIZATION TIME OF PIG OOCYTES MATURED IN-VITRO BY BOAR SPERM STORED AT 4°C

2004 ◽  
Vol 16 (2) ◽  
pp. 264
Author(s):  
Y.J. Yi ◽  
M.Y. Kim ◽  
Y.J. Chang ◽  
D.I. Jin ◽  
C.S. Park
Keyword(s):  
In Vitro Fertilization ◽  
Room Temperature ◽  
Sperm Concentration ◽  
Egg Yolk ◽  
Penicillin G ◽  
In Vitro Production ◽  
Vitro Fertilization ◽  
Boar Sperm ◽  
Sperm Penetration

The use of boar sperm stored at 4°C may be a useful tool for in vitro production of pig embryos. Therefore, this study was undertaken to investigate the effect of fertilization time of pig oocytes matured in-vitro by boar sperm. The sperm-rich fraction (30–60mL) was slowly cooled to room temperature (20–23°C) by 2h after collection. Semen was transferred into 15-mL tubes, centrifuged at room temperature for 10min at 800g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5mL of the LEN (11.0g lactose hydrate, 20mL egg yolk, 0.05g N-acetyl-D-glucosamine and 100mL distilled water) diluent to provide 1.0×109 spermmL−1 at room temperature. The resuspended semen was cooled in a refrigerator to 4°C. The medium used for oocyte maturation was TCM-199 supplemented with 26.19mM sodium bicarbonate, 0.9mM sodium pyruvate, 10μgmL−1 insulin, 2μgmL−1 vitamin B12, 25mM HEPES, 10μgmL−1 bovine apotransferrin, 150μM cysteamine, 10IUmL−1 PMSG, 10IUmL−1 hCG, 10ngmL−1 EGF, 0.4% BSA, 75μgmL−1 sodium penicillin G, 50μgmL−1 streptomycin sulfate and 10% pFF. After about 22h of maturation, oocytes were cultured without cysteamine and hormones for 22h at 38.5°C, 5% CO2 in air. Oocytes were inseminated with boar sperm stored at 4°C for 2 days after collection. Oocytes were coincubated for 1, 3, 6 and 9h in 500μL TBM fertilization media with 1×106mL−1 sperm concentration. Thereafter, oocytes were transferred into 500μL NCSU-23 culture medium containing 0.4% BSA for further culture of 6, 48 and 144h, fixed and stained for the evaluation of fertilization parameters and developmental ability. Data were analysed by ANOVA and Duncan’s multiple range test using the SAS program. The rates of sperm penetration and male pronuclear formation were higher in the fertilization times of 6 and 9h than in those of 1 and 3h. The percentage of polyspermic oocytes was highest in fertilization time of 9h compared with other incubation times. The rates of cleaved oocytes were higher in the fertilization times of 6 and 9h (85.0 and 84.6%) compared with those of 1 and 3h (61.1 and 76.8%). The percentage of blastocyst formation from the cleaved oocytes was highest in the fertilization time of 6h (33.6%) than in that of 1, 3 and 9h (11.4, 23.0 and 29.6%). Mean cell numbers per blastocyst were 32.9±3.3, 27.6±2.7, 26.3±2.2 and 24.4±1.8 in the fertilization times of 6, 9, 3 and 1h, respectively. In conclusion, we found out that boar sperm stored at 4°C could be used for in vitro fertilization of pig oocytes matured in vitro. Also, we recommend the coincubation time of 6h in 500μL TBM fertilization medium with 1×106mL−1 sperm concentration for in vitro fertilization of pig oocytes matured in vitro.

scholarly journals Improvement of in vitro fertilization by a tannin rich vegetal extract addition to frozen thawed boar sperm

2020 ◽  
Vol 17 (2) ◽  
Author(s):  
Giovanna Galeati ◽  
Diego Bucci ◽  
Chiara Nerozzi ◽  
Beatrice Gadani ◽  
Carlo Tamanini ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Vitro Fertilization ◽  
Boar Sperm

scholarly journals 304 PRODUCTION OF PORCINE EMBRYOS OF A PREDETERMINED SEX AFTER IN VITRO FERTILIZATION OF IN VITRO-MATURED OOCYTES WITH SEX-SORTED FROZEN-THAWED BOAR SPERM

2005 ◽  
Vol 17 (2) ◽  
pp. 303 ◽  
Author(s):  
R. Bathgate ◽  
K.M. Morton ◽  
B.M. Eriksson ◽  
D. Rath ◽  
B. Seig ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Restriction Analysis ◽  
Motile Sperm ◽  
Large White ◽  
Vitro Fertilization ◽  
Boar Sperm ◽  
Fort Collins ◽  
Cattle And Sheep

Pre-sexed embryos and offspring have been produced after IVF and embryo transfer (ET) with sex-sorted frozen-thawed sperm in cattle and sheep (Maxwell et al. 2004 Anim. Reprod. Sci. 82–83, 79–95). The aims of this study were to demonstrate that sex-sorted frozen-thawed boar sperm could be incorporated into pig IVF for the production of embryos of a predetermined sex and that these embryos could be successfully nonsurgically transferred. Ovaries were collected from abattoir slaughtered gilts (n = 138) and selected COCs were matured in vitro (Long et al. 1999 Theriogenology 51, 1375–1390). Sperm were collected from a mature boar and diluted with Androhep (1:3, semen:Androhep; Minitube, Verona, WI, USA), stained with H33342, and separated into X and Y sperm using a SX MoFlo (Cytomation, Inc., Fort Collins, CO, USA). Sex-sorted sperm were cryopreserved in 0.5 mL straws using the Westendorf protocol modified for sorted sperm (Bathgate, unpublished). Thawed sperm (Y sperm only) were prepared for IVF by centrifugation (300g, 10 min) through a Porcipure gradient (Nidacon Int. AB, Gothenburg, Sweden), and washed (centrifugation 300g, 10 min) in mTALP-PVA. For IVF, COCs were denuded and groups of 100 oocytes were transferred to 200-μL drops of mTALP-PVA (Long et al. 1999) and incubated with 5,000 motile sperm for 4–6 (Short) or 18–20 h (Long) . Presumptive zygotes were washed and transferred to 100-μL drops of mNCSU-23 (Long et al. 1999) and cultured until Day 4 (Day 0 = IVF) in humidified 5% CO2, 6% O2, 89% N2. Oocyte cleavage was assessed 48 h post-insemination, and on Day 4 selected morulae were transferred to recipient sows (n = 7 Large White × Landrace; 65 morulae per sow) nonsurgically using a Firflex catheter (Magapor, Zaragoza, Spain). Sex of remaining embryos was confirmed by PCR and restriction analysis (Cong et al. 1993 Hum. Mol. Genet. 2 1187–1191). Data from three replicates were arc sin transformed and analyzed by ANOVA. Oocyte cleavage was similar after Short (724/1547; 46.8%) or Long (598/1528; 39.1%) co-incubation. Resort analysis showed sperm to be >91% purity, and all sexed morulae were of the predicted sex (16/16). Delayed return to estrus (>23 days) was observed in 5 recipient sows (71.4%). Fetal sacs were observed by transcutaneous ultrasound in one of these sows. Pre-sexed porcine IVP embryos can be successfully produced using sex-sorted frozen-thawed boar sperm, and these embryos are capable of initiating pregnancies when transferred to recipients. However, further refinement of porcine IVP and ET protocols are required to enable full in vivo development. This work was supported by XY, Inc., Fort Collins, CO, USA.

275 PENTOXIFYLLINE ADDED TO FREEZING EXTENDER HAS A DELETERIOUS EFFECT ON POST-THAW BOAR SPERM QUALITY AND IN VITRO FERTILIZATION RATE

2006 ◽  
Vol 18 (2) ◽  
pp. 245
Author(s):  
M. A. Gil ◽  
J. Roca ◽  
M. Hernandez ◽  
C. Cuello ◽  
C. Almiñana ◽  
...  
Keyword(s):  
In Vitro Fertilization ◽  
Deleterious Effect ◽  
Sperm Quality ◽  
Membrane Integrity ◽  
Control Group ◽  
Vitro Fertilization ◽  
Split Plot ◽  
Boar Sperm ◽  
Plot Design

Pentoxifylline, a methylxanthine derivative, is considered to be a hyperactivation and acrosome reaction-improving agent. The purpose of this study was to test how the addition of pentoxifylline to freezing extender influences post-thaw sperm motility and membrane integrity. The ability of thawed spermatozoa to fertilize in vitro-matured oocytes was also assessed. Pooled ejaculate sperm-rich fractions collected from three fertile boars were frozen in 0.5-mL straws after being diluted in lactose/egg yolk/glycerol/Orvus-ES-Paste extender (0 pentoxifylline = control) or the same extender supplemented with 2, 4, 8, 16, and 32 mM pentoxifylline. To evaluate post-thaw sperm survival, the percentage of total motile spermatozoa and rapid progressive spermatozoa (CASA system) and plasma membrane and acrosome integrity (flow cytometry) were assessed. Data from six replicates were analyzed in a split plot design using a PROMIXED model. The addition of 4, 8, 16, and 32 mM pentoxifylline to freezing extender significantly decreased progressive and total motility (P < 0.001) compared with control (4.5/26.6%, 4.5/24.5%, 2.8/20.5%, 0.6/11.4%, and 13.2/49.7% for the 4, 8, 16, and 32 mM pentoxifylline groups and the control group, respectively). The same was observed for sperm viability; the percentage of viable spermatozoa with intact acrosomes was significantly lower (P < 0.001) in pentoxifylline-treated groups compared with the control group, chiefly in the 16 mm, and 32 mM pentoxifylline groups (54, 11.6, and 6.2% for control, 16 and 32 mM, respectively). To evaluate in vitro fertilization parameters, cumulus-oocyte complexes were matured in BSA-free NCSU23 medium with 10% porcine follicular fluid, 0.1% cysteine, 10 ng EGF, 10 IU/mL eCG, and 10 IU/mL hCG, in a incubator at 39�C and 5% CO2. After 40-44 h of maturation, oocytes (n = 1067, in three replicates) were denuded of cumulus cells, washed, transferred to droplets (30 oocytes in 50 �L) of TBM medium supplemented with 2 mM caffeine and 0.2% BSA, and inseminated (2000 thawed sperm/oocyte). After a co-incubation period of 6 h, oocytes were washed and transferred to droplets (500 �L) of NCSU23 with 0.4% BSA for another 10-14 h, then fixed and stained for 72 h, and examined under a phase-contrast microscope. Data were analyzed as split plot design using a PROMIXED model. The addition of pentoxifylline to the freezing extender reduced significantly (P < 0.001) the penetration rate (51.4, 17.5, 15.8, 17.8, 9.5, and 4.8% for the 0, 2, 4, 8, 16, and 32 mM pentoxifylline groups, respectively) and the efficiency (monospermic oocytes/total inseminated oocytes) of fertilization (34.8, 14.6, 14.7, 15.5, 7.9, and 4.8% for the 0, 2, 4, 8, 16, and 32 mM pentoxifylline groups, respectively) as compared with the control group (the first value in each of these two cases). It is therefore concluded that pentoxifylline added to the freezing extender has a deleterious effect on post-thaw boar sperm quality and on their ability to fertilize the oocytes in vitro. This work was supported by CICYT (AGL05-0471).

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