• Journal of Embryo Transfer
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• v.15 no.3
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• pp.237-245
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• 2000

This study was carried out to elucidate whether sperm contain a factor inducing second polar body extrusion and to search for an effective collection method of the sperm factor Thus, sperm extract, dialyzed sperm-extract or liquid chromatographic fractions of sperm extract was microinjected into ovulated oocytes. And the microinjected oocytes were incubated for 24 hours to investigate about the extrusion of second polar body. The results obtained were as follows; 1. Sperm extract significantly increased the second polar body extrusion. 2. Sperm extract showed five major fractions at retention volumes (RVs) 1.25, 1.37, 1.84, 2.10 and 2.67ml after separation with Superose 12 column. These sperm extract fractions did not significantly increase the second polar body extrusion. 3. Dialyzed sperm-extract significantly increased the second polar body extrusion 4. Dialyzed sperm-extract showed three maior fractions at RVs 1.88, 2.14 and 2.77ml after separation with Superose 12 column. Of these fractions, the fraction RV2.14 significantly increased the second polar body extrusion. In conclusion, sperm extract contained a factor inducing the second polar body extrusion and the factor was contained largely in fraction RV2.14 after dialysis and liquid chromatographic fractionation of sperm extract.

• 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.

• Korean Journal of Poultry Science
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• v.33 no.2
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• pp.165-169
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• 2006

The objective of this study was to elucidate the basis for the difference in sperm production of broiler breeders. Nine sexually mature Hubbard broiler breeder males, 35 weeks of age, were trained for two weeks for semen collection on alternate days by abdominal massage technique. Following the training, the breeder males were collected daily for five successive days. The males were then classified as low or high sperm producers. The mean body weights of individual males were also recorded on the basis of body weight at the start and end of the experiment. Immediately after last collection the males were slaughtered and testes biometry was determined. Daily sperm output of individual males varied from $0.21{\times}10^9\;to\;2.64{\times}10^9$ sperm. The daily sperm production of low sperm producer males was lower ($0.47{\pm}0.13\;vs.\;2.06{\pm}0.20{\times}10^9$; P<0.05) than high sperm producer males. Testes weight of low sperm producer males was lower ($6.32{\pm}1.6\;vs.\;20.33{\pm}4.76\;gm$; P<0.05) than high sperm producer males. Moreover the testis weight of high sperm producer males was 3.22 times higher than low sperm producer males. The average body weight of high sperm producer males was higher ($4,389{\pm}116.3\;vs.\;3,960{\pm}131.77\;gm$; P>0.05) than low sperm producer males. The correlation coefficients indicate significantly positive correlation of body weight (P<0.05) and testes weight (P<0.01) on semen volume, sperm concentration and daily sperm production.

• Animal Bioscience
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• v.34 no.8
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• pp.1253-1270
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• 2021

Assessment of male fertility is based on the evaluation of sperm. Semen evaluation measures various sperm quality parameters as fertility indicators. However, semen evaluation has limitations, and it requires the advancement and application of strict quality control methods to interpret the results. This article reviews the recent advances in evaluating various sperm-specific quality characteristics and methodologies, with the help of different assays to assess sperm-fertility status. Sperm evaluation methods that include conventional microscopic methods, computer-assisted sperm analyzers (CASA), and flow cytometric analysis, provide precise information related to sperm morphology and function. Moreover, profiling fertility-related biomarkers in sperm or seminal plasma can be helpful in predicting fertility. Identification of different sperm proteins and diagnosis of DNA damage has positively contributed to the existing pool of knowledge about sperm physiology and molecular anomalies associated with different infertility issues in males. Advances in methods and sperm-specific evaluation has subsequently resulted in a better understanding of sperm biology that has improved the diagnosis and clinical management of male factor infertility. Accurate sperm evaluation is of paramount importance in the application of artificial insemination and assisted reproductive technology. However, no single test can precisely determine fertility; the selection of an appropriate test or a set of tests and parameters is required to accurately determine the fertility of specific animal species. Therefore, a need to further calibrate the CASA and advance the gene expression tests is recommended for faster and field-level applications.

• Proceedings of the KSAR Conference
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• 2003.06a
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• pp.63-63
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• 2003

This study was carried out to investigate the effects of liquid boar sperm stored at 4$^{\circ}C$ on sperm motility, normal acrosome, and in-vitro fertilization and culture of pig oocytes matured in-vitro. The sperm-rich fraction (30~60 ml) of ejaculate was collected into an insulated vacuum bottle. Semen was slowly cooled to room temperature (20~23$^{\circ}C$) by 2 h after collection. Semen was transferred into 15 ml tubes, centrifuged at room temperature for 10 min at 800$\times$g, and the supernatant solution was poured off. The concentrated sperm was resuspended with 5 ml of lactose, egg yolk and N-acetyl-D-glucosamine (LEN) diluent to provide 1.0$\times$10$^{9}$ sperm/ml at room temperature. The resuspended semen was cooled in a refrigerator to 4$^{\circ}C$ and preserved for 5 days to examine sperm motility and normal acrosome. The medium used for oocyte maturation was modified tissue culture medium (TCM) 199. After about 22 h of culture, oocytes were cultured without cysteamine and hormones for 22 h at 38.5$^{\circ}C$, 5% $CO_2$ in air. Oocytes were inseminated with liquid boar sperm stored at 4$^{\circ}C$ for 2 days after collection. Oocytes were coincubated for 6 h in 500 ${mu}ell$ mTBM fertilization media with 0.2, 1, 5 and 10$\times$10$^{6}$ /ml sperm concentration, respectively. At 6 h after IVF, oocytes were transferred into 500 ${mu}ell$ Hepes-buffered NCSU-23 culture medium for further culture of 6, 48 and 144 h. There were significant differences in sperm motility and normal acrosome among preservation days and incubation times, respectively. The rates of sperm penetration and polyspermy were higher in 5 and 10$\times$10$^{6}$ sperm/ml than in 0.2 and 1$\times$10$^{6}$ sperm/ml. Male pronuclear formation was lower in 0.2$\times$10$^{6}$ sperm/ml than in 1, 5 and 10$\times$10$^{6}$ sperm/ml. Mean numbers of sperm in penetrated oocyte were highest in 10$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. The rate of blastocysts from the cleaved oocytes (2~4 cell stage) was highest in 1$\times$10$^{6}$ sperm/ml compared with other sperm concentrations. In conclusion, we found out that liquid boar sperm stored at 4$^{\circ}C$ could be used for in-vitro fertilization of pig oocytes matured in-vitro. Also, we recommend 1$\times$10$^{6}$ ml sperm concentration for in-vitro fertilization of pig oocytes.

• Journal of Embryo Transfer
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• v.26 no.4
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• pp.287-296
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• 2011

Mammalian fertilization is a complex cascade process consisting of sperm migration through the female reproductive tract, physiological changes to sperm such as sperm capacitation and acrosome reaction, and sperm-egg interaction in the oviduct in vivo. On the other hand, in vitro fertilization (IVF) is a process by which egg cells are fertilized by sperm outside the body: in vitro. IVF has been used for a variety of purposes in reproductive biotechnology for human and animals. The discovery of sperm capacitation in 1951 promoted the development of IVF technology. In the initial stage of IVF, sperm capacitation in preincubation medium was shown to be essential to fuse with eggs. Besides, sperms should detour some of the in vivo regulations for IVF. This review introduces a general mammalian fertilization process, including sperm capacitation, removal of cumulus matrix, acrosome reaction, and sperm-egg fusion and focuses on the roles of key biochemical molecules, signal mechanisms, and genes involved during IVF and novel results of sperm-oocyte interaction elucidated in various gene-knockout mice models.

• Journal of Embryo Transfer
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• v.9 no.3
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• pp.269-277
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• 1994

The objective of this study was to develop an effective in vitro production system capable of obtaining more porcine embryos from immature oocytes These experiments were conducted to examine the effect of sperm factor on the IVF and IVD, and the effect of coculture with somatic cells on the IVD of embryos. Although the concentration of epididymal sperm for IVF did not affect on cleavage rate, but 5 x 105 sperm/mi showed the highest cleavage rate(48.7%) and the developmental potential of IVF oocytes from this concentration was also greatly higher (P$^{\circ}C$-stored sperm for l2hrs and the cleavage rate from fresh sperm was significantly higher (P<0.05) than that from frozen sperm, but the developmental potential after IVF was slightly high from the frozen sperm. The cleavage rate of IVF oocytes cocultured with oviductal epithelial cells and cumulus cells was 76.3% and 72.9%, respectively. There was no difference between two coculture systems but this rate was significantly higher(P<0.05) than that of medium alone(42.0%).

• Journal of Veterinary Clinics
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• v.18 no.4
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• pp.345-349
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• 2001

The aim of this study was to investigate the effect of sperm preservation according to the various kinds of commercially available semen extenders and to investigate the effect of sperm preservation according to the various temperature storages of fresh-extended porcine semen. To investigate the effect of sperm preservation according to the various kinds of commercially available semen extenders, porcine semens diluted in 3 semen extenders, Beltsville Thawing Solution(BTS), Androhep and Kiev, were cooled at $8^{\circ}C$ storage temperature with a controlled cooling rate of $2-4^{\circ}C$/h. Motility, progressive motility, normal sperm(%) and sperm morphology were assessed comparatively. In motility and progressive motility, Androhep extenders revealed better result than other extenders. In normal sperm(%) and sperm morphology, 3 semen extenders revealed similar results. To investigate the effect of sperm preservation according to the various temperature storages of fresh-extended porcine semen, porcine semens diluted in BTS extender, were cooled at 3 storage temperatures($8^{\circ}C$, $12^{\circ}C$ and $17^{\circ}C$) with a controlled cooling rate of $2-4^{\circ}C$/h. Motility, progressive motility, normal sperm(%) and sperm morphology were assessed comparatively. In motility and progressive motility, $8^{\circ}C$ treatment group revealed better result than $12^{\circ}C$ and $17^{\circ}C$ treatment groups. In normal sperm(%) and sperm morphology, 3 temperatures of treatment groups revealed similar results.

• Journal of Animal Reproduction and Biotechnology
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• v.36 no.4
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• pp.239-246
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• 2021

The immunological sperm separation method is economical compared to the existing sorting method, and it is promising for the development of new technologies by reducing sperm damage. Wholemom (WM) is a sex-regulating protein that comprises on immunoglobulin G coupled with magnetic nanoparticles (MNPs) that responds to surface proteins derived from the Y chromosome in cattle. Y sperms are restricted in motility as the WM aggregates them, and the magnet could separate the non-aggregated cells. This study was conducted to investigate the effect of WM treatment on the characteristics of bull sperm. After treating sperm with WM and incubation for 6 h, the motility parameters including total motility, progressive motility, velocity average path, velocity straight line, amplitude of lateral head displacement, and linearity were significantly higher in the WM treatment group than in the control group (p < 0.05). Sperm viability and acrosome reaction rates were similar in both groups during each incubation period (p > 0.05). In conclusion, the immunological sperm sexing procedure using a monoclonal antibody conjugated with MNPs did not affect the characteristics of bull sperm. This study suggests that compared to other techniques, the immunological method for sperm sexing could classify sperm quickly and efficiently without the use of expensive equipment.

• Korean Journal of Veterinary Research
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• v.32 no.3
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• pp.435-450
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• 1992

To evaluate the fertilizing capacity of domestic animal spermatozoa by hamster test, semen were collected from 15 boars(Duroc, Landrace, and Yorkshire) and 2 mixed dogs which had been proved to be fertile in the past then, the semen were preserved in BWW medium at $4^{\circ}C$ or $18^{\circ}C$ for about 20 hours and coincubated with zona-free hamster ova for 5 hours. The ova were stained by lacmoid and examined under phase contrast microscope to investigate the rates of sperm binding to the ova, penetration and formation of a male pronucleus, and the numbers of both bound and penetrated sperm per ovum. Both the semen preserved at $18^{\circ}C$ for about 20 hours and that treated by swim up procedure showed considerably higher rates of sperm binding and penetration as well as higher number of penetrated sperm than that preserved at $4^{\circ}C$ for about 20 hours, respectively(p<0.01). Motility of boar sperm at insemination was from 40 to 90% and no difference in hamster test was obtained according to different degree of sperm motility. Abnormality in morphology of boar sperm at insemination was from 6 to 45% and no difference in hamster test was obtained according to different degree of sperm abnormality. The sperm concentrations of $7{\times}10^7$ and $7{\times}10^6$ showed considerably higher rates of sperm binding and penetration as well as higher number of bound sperm than that of $7{\times}10^4$ (p<0.01) along with the same higher results than that of $7{\times}10^5$(0<0.05), respectively. Boar sperm showed considerably higher rates of sperm binding and penetration as well as higher numbers of both bound and penetrated sperm than dog sperm, when both semen were treated by BWW＋heparin medium and swim up procedure, respectively. These results indicated that fertile boar sperm showed considerably lower rates in the results of hamster test, when preserved at $4^{\circ}C$ for about 20 hours and in lower concentration of sperm than when preserved at $18^{\circ}C$ for about 20 hours and in higher concentration of sperm, respectively, and at the same time considerably higher results than fertile dog sperm, consequently to prove that hamster test would be of great value in assaying the fertilizing capacity of boar sperm.