• Journal of Embryo Transfer
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• v.14 no.1
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• pp.59-67
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• 1999

To make up the medium for quantitative selection of capacitated-sperm through sucrose layer, the effects of BSA, caffeine, heparin and progesterone on sperm swim-up migration and movement were examined. And the results obtained were as follows; 1. BSA of 4mg/ml in bMSS stimulated sperm migration and movement, and attracted capacitated-sperm. 2. Caffeine of 5mM in bMSS containing 4mg/ml BSA stimulated sperm movement and attracted capacitated-sperm. 3. Heparin of 20$\mu\textrm{g}$/ml in bMAA containing both 4mg/ml BSA and 5mM caffeine stimulated movement and capacitation of sperm. 4. Progesterone of 50$\mu\textrm{g}$/ml in bMSS containing all 4mg/ml BSA, 5mM caffeine and 20$\mu\textrm{g}$/ml heparin (BCHP-MSS) attracted capacitated-sperm. 5. Effect of BCHP-MSS on sperm on sperm attraction was not different from effect of 10% follicular fluid solution (FF-MSS) on sperm swim-up separation. In conclusion, bMSS with 4mg/ml BSA, 5mM caffeine, 20$\mu\textrm{g}$/ml heparin and 50$\mu\textrm{g}$/ml progesterone(BCHP-MSS) was a optimal condition for selection of capacitated-sperm through sucrose layer.

• Journal of Embryo Transfer
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• v.11 no.3
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• pp.309-316
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• 1996

The aim of this study is to elucidate sperm chemotaxis and to set up the optirnal condition for selection of motile and capacitated sperm from hovine frozen-semen. Thus, the effects of semen-washing after thawing, concentrations of progesterone (P4) and bovine serum albumin (BSA), and sperm-washing frequency on sperm selection were examined. For evaluating their effects, number, viability and acrosome reaction of sperm swim-up seperated from semen, which were incubated for 30 minutes at 36$^{\circ}C$ in the M2 solution containing P4 and BSA, were investigated. For frozen-semen just after thawing, sperm recovery and viability were not significantly different between P4-treated and -untreated semen. However, washing frozen-semen decreased the number of sperm and increased the viability of sperm that were recovered from semen treated with P4. Progesterone affected the recovery rate, the viability and the acrosome-reaction rate of sperm recovered from washed frozen-semen. Especially, number of motile and capacitated sperm were highest in semen treated with 50$\mu$g /ml among 0, 20, 50 and 100$\mu$g /ml of P4 concentrations. BSA affected the recovery rate and the viability of sperm recovered from washed frozen-semen that were treated with 50$\mu$g /ml of P4. Especially, the percentage of viable sperm were highest in semen treated with 4mg /ml among 0, 2, 4, and 6mg /ml of BSA concentrations. Repeatedly sperm-washing did not affect the recovery rate and the viability of sperm recovered from washed frozen-semen that were treated with 50$\mu$g /ml of P4 and 4mg /ml of BSA In conclusion, using progesterone and BSA could efficiently make the selection of motile and capacitated sperm from washed frozen-semen.

• Journal of Aquaculture
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• v.19 no.1
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• pp.52-56
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• 2006

This study was conducted to investigate the effects of three different concentrations (6%, 8% and 10% final volume) of dimethyl sulfoxide (DMSO) on cryopreserved sperm of grass carp (Ctenopharyngodon idellus). Grass carp sperm was suspended in Kurokura extender #2 and equilibrated at $4^{\circ}C$ for 10 min. French straws (0.25 ml) of sperm were frozen from $4^{\circ}C\;to\;-4^{\circ}C$ at a rate of $4^{\circ}C\;min^{-1}$ and then ken $-4^{\circ}C\;to\;-80^{\circ}C$ at a rate of $11^{\circ}C\;min^{-1}$. The straws were kept at $-80^{\circ}C$ for 10 min and finally stored in liquid nitrogen $(-196^{\circ}C)$. The cryopreserved sperm was thawed in a water bath at $40^{\circ}C$ for 30 sec and fertilization, hatching rate and larval malformation were compared with fresh sperm (control). The fertilization rate of post-thawed sperm was comparable (from 88.21% to 94.30%) to that of fresh sperm. However, hatching rate of all frozen sperm were significantly lower (P<0.05) than that of control. Additionally, the larval abnormality rate of frozen sperm was significantly higher than that of fresh sperm. The results indicate that DMSO could affect the quality of cryopreserved sperm of grass carp, and a freezing program and a proper extender composition should be further studied.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.26-26
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• 2003

Hyaluronic acid (HA)-binding sites have been shown the diagnostic potential fur assessment of sperm maturity, which is related to male fertility. This study was designed to evaluate chromosomal patterns in porcine embryos produced by in vitro fertilization (IVF) and intracytoplasmic sperm injection (ICSI) with non- or HA-binding sperm (HABS). For binding of sperm with HA, sperm incubated in 10 ${mu}ell$ drop containing HA (0.8 mg/ml)-agarose (0.8%) mixture for 15 min. IVF and ICSI with non- or HA-bound sperm examined with matured oocytes at 44 hr after in vitro maturation. Embryos were cultured in 50 ${mu}ell$ of NCSU 23 containing 0.5% BSA for 5 days and then in 50 ${mu}ell$ of NCSU 23 containing 10% FBS for 2 days. For the evaluation of chromosomal aneuploidies, chromosome 1 sub-metacentric specific probe was used in sperm and embryos by fluorescence in situ hybridization (FISH). The frequency of aneuploidy sperm for chromosome 1 was 6.25%. The significant differences following IVF and ICSI with non- or HA-bound sperm were not observed in blastocyst formation rates (18.6, 23.5, and 23.8%) and cell number (61.8 $\pm$ 12.5, 55.5 $\pm$ 7.3, and 59.3 $\pm$ 9.6). Moreover, the percentage of diploidy in 4-cell stage embryos was 57.1% (IVF), 68.8% (ICSI), and 76.3% (ICSI-HABS). These results suggest that HA-binding sites may be a material for selection of normal sperm for ICSI. Therefore HA selection of normal sperm may be reduce the loss to embryonic mortality prior to embryo transfer in pig.

• Korean Journal of Veterinary Research
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• v.29 no.1
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• pp.57-60
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• 1989

This study was carried out to evaluate the possibility of sex preselection by gradients methods using bovine serum albumin in rabbits. Artificial insemination was performed with sperm from the top and bottom layer of rabbit semen separated by bovine serum albumin gradients. Various characteristics of separated sperm, and the conception rate and secondary sex ratio at artificial insemination with separated sperm were compared. The results obtained were as follows. 1. The sperm from the bottom layer showed significanty high value in motility, percent of normal sperm and progressive motility as compared with control sperm and the sperm and the sperm from the top layer. 2. The conception rate of sperm from the bottom layer was higher than that of the top layer. But secondary sex ratio was not altered by this methods.

• Journal of Embryo Transfer
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• v.7 no.2
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• pp.133-136
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• 1992

Glass wool filtration and swim-up method resulted in inreasing to 58.3% and 62.7% of the progressive motility in frozen-thawed boar sperm, compared to 34.2% in the untreated sperm. Glass wool filtration tended to be more successful than swim-up method for the survival sfter incubation of 38.5$^{\circ}C$ for 3h. Sperm recovered by both the swim-up method and the glass wool filtration method were tested in an in vitro fertilization to determine which of the two techniques would yield sperm with high fertilizing capacity. The results indicated that there was a significantly(p

• Asian-Australasian Journal of Animal Sciences
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• v.27 no.10
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• pp.1417-1425
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• 2014

The selection of morphologically normal spermatozoa is critical to obtain high breeding performances in boar breeding farms and artificial insemination (AI) centers. Parameters for the selection of semen mainly include total sperm motility, concentration, and morphology. However, these primary parameters are often not reliable for discriminating between normal and abnormal, non-fertilizable spermatozoa. The present study was designed to compare the motion characteristics, fertilization ability using in vitro fertilization (IVF), and acrosome formation of the semen from boars having low (boar number 2012) and normal (boar number 2004 and 2023) breeding performances. The ultimate goal was to identify additional simple and easy criteria for the selection of normal sperm. There was no significant difference between boar 2004 and boar 2023 sperm total motility in computer assisted sperm analysis. However, boar number 2012 semen presented a significantly reduced population of rapid moving spermatozoa and an increased population of slow moving spermatozoa compared to boar numbers 2004 and 2023. Analysis of detailed motion characteristics revealed that sperm from boar number 2012 had significantly reduced motility in progressiveness, average path velocity, straight-line velocity (VSL), curvilinear velocity (VCL), straightness, and linearity. The assessment of the fertilizing ability by IVF also showed that sperm from boar number 2012 showed a fertility rate of 3.4%, whereas sperm from boar number 2023 had a fertility rate of 75.45%. Interestingly, most of the sperm nuclei were found on the peripheral area of the oocytes, suggesting that the sperm from boar number 2012 lacked penetration ability into the oocyte zonapellucida. The acrosome formation analysis using Pisum sativum agglutinin staining demonstrated that the sperm from boar number 2012 had a defect in acrosome formation. Consequently, primary parameters for selecting semen before AI such as motility are not sufficient to select normal and fertilizable spermatozoa. In conclusion, the present study suggests that the acrosome staining and detailed motion characteristics such as progressiveness, VCL, and VSL should be included in determining semen quality together with primary parameters for successful AI and high breeding performance in the swine industry.

• Clinical and Experimental Reproductive Medicine
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• v.45 no.3
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• pp.101-109
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• 2018

In an age when a small quantity of sperm can lead to pregnancy through in vitro fertilization or intracytoplasmic sperm injection, selecting healthy sperm is important. Sperm DNA fragmentation (SDF) is known to be higher in infertile men. Terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling (TUNEL) and the alkaline comet test are SDF tests that directly measure DNA damage and have shown closer correlations with assisted reproduction results than indirect tools such as the sperm chromatin structure assay or the sperm chromatic dispersion test. It is difficult; however, to endorse a single test as the best test overall; instead, it is best to select a testing method based on each patient's clinical condition and goals. In a couple struggling with infertility, if the male partner has a high level of SDF, he should aim to decrease SDF through lifestyle modifications, antioxidant treatment, and ensuring an appropriate duration of abstinence, and physicians need to treat the underlying diseases of such patients. If sperm DNA damage continues despite the patient's and physician's efforts, other methods, such as micromanipulation-based sperm selection or testicular sperm extraction, should be used to select healthy sperm with nuclear DNA integrity.

• Asian-Australasian Journal of Animal Sciences
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• v.31 no.9
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• pp.1387-1392
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• 2018

Objective: In a nucleus breeding scheme, the sire of dam's pathway plays an important role in producing genetic improvement. Selection proportion is the key parameter for predicting selection intensity, through truncating the normal distribution. Semen sexing using flow cytometry reduces the number of vials of sperm that can be obtained from a proved bull. In addition, a lower fertility of this kind of sperm is expected because of the lower sperm dosage in sex sorted semen. Both of these factors could affect the selection proportion in the sire of dam's pathway ($p_{SD}$). Methods: In the current study, through a deterministic simulation, effect of utilizing sex sorted semen on selection ($p_{SD}$) was investigated in three different strategies including 1: continuous use of sex sorted semen in heifers (CS), 2: the use of sex sorted semen for the first two (S2) and 3: the first (S1) inseminations followed by conventional semen. Results: Results indicated that the use of sex sorted semen has a negative impact on the sire of dams (SD) pathway due to increase in selection proportion. Consequently selection intensity was decreased by 10.24 to 20.57, 6.38 to 8.87 and 3.76 to 6.25 percent in the CS, S2, and S1 strategies, respectively. Conclusion: Considering the low effect of sexed semen on genetic improvement in dam pathways, it is necessary to consider the joint effect of using sex sorted semen on the sire and dams pathway to estimate about the real effect of sexed semen on genetic improvement in a nucleus breeding scheme.

• Journal of Animal Reproduction and Biotechnology
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• v.38 no.1
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• pp.38-46
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• 2023

In this study, an experiment was conducted in order to determine what cryopreservatives (CPVs) were more effective in supporting the motility and viability of sperm from experimental animals. The sperm of mice, rats, beagle dogs, and rabbits were frozen using different CPVs, including DMSO, TYB, and Sperm CryoProtec. The results from freezing the sperm of each laboratory animal in Sperm CryoProtec showed a high level of sperm motility and viability in sperm samples from mice, rats, and beagle dogs melted at the end of the first week. For rabbits, a high level of motility was observed in sperm thawed during the first week, whereas a high level of viability was observed in sperm thawed during the second week. The results of analysis of sperm motility and viability using different CPVs according to laboratory animals showed a significantly higher level of sperm motility (26.28%) and viability (36.20%) for mice in Sperm CryoProtec and the lowest levels of motility and viability were observed in DMSO (p < 0.05). Significantly higher levels of motility (27.94%) and viability (37.94%) were observed for rats in Sperm CryoProtec compared with TYB, which showed the lowest levels of motility and viability (p < 0.05). The study findings described above suggest that the selection of appropriate cryopreservatives is required for each experimental animal. This is because there are differences in the levels of sperm motility and viability of experimental animals depending on the CPVs that are typically used for freezing human sperm, including Sperm CryoProtec and TYB.