• Proceedings of the KSAR Conference
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• 2001.10a
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• pp.56-56
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• 2001

This study was carried out to investigate on the improvement of fertilizing and developing ability of in vitro matured oocytes from individuals of bulls, sperm type, pretreatment of sperm or oocytes obtained by intracytoplasmic sperm injection(ICSI). 1. The male pronuclear formation and developmental rates of oocytes obtained by ICSI treated individual of bulls were 73.9%-87.0% and 33.3%-60.9%, respectively. 2. The male pronuclear formation and developmental rates of oocytes obtained by ICSI treated fresh and frozen sperm, tail-cutting and tail-scoring sperm were 82.0%, 78.0%, 42.2%, 51.1% and 56.0%, 42.0%, 17.8%, 22.2% respectively. and these values of fresh sperm injection were higher than that of frozen sperm, tail-cutting and tail-scoring. 3. The male pronuclear formation and developmental rates of oocytes obtained by sperm pretreated heparin, BFF(bovine follicula fluid), His, Ca Ionophore(Ⅰ) and Ⅰ ＋ caffeine methods were 66.7%-82.2% and 33.3%-60.6%, respectively. and these values of treatment of Ⅰ＋ caffeine were higher than that of other methods. 4. The male pronuclear formation and developmental rates of oocytes obtained by ICSI treated with or without zona pellucida were 80.0%, 72.0% and 46.0%, 36.0%, respectively.

• 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

• Journal of Microbiology and Biotechnology
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• v.13 no.6
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• pp.833-840
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• 2003

Sperm motility of salmonid fishes is suppressed by external $K^{+}$ and initiated by decrease of $K^{+}$ concentration surrounding the sperm. It was shown that the decrease in external $K^{+}$ concentration induced not only the initiation of sperm motility, but also hyperpolarization of the plasma membrane and synthesis of cAMP in the sperm of rainbow trout, steelhead trout, and masu salmon. Inhibitors of $K^{+}$ channels, especially voltage-dependent $K^{+}$ channels, inhibited these three reactions, and the inhibitions were abolished by subsequent addition of a $K^{+}$ ionophore, valinomycin, suggesting that $K^{+}$ efflux through the $K^{+}$ channel contributes to rapid changes in the membrane potential of sperm and cAMP synthesis, thereby resulting in the initiation of sperm motility of salmonid fishes.

• Clinical and Experimental Reproductive Medicine
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• v.18 no.1
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• pp.89-94
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• 1991

Thirty five couples were treated by intrauterine insemination with sperm prepared by a washing and swim-up method. Fifteen women conceived(42.9%). Sperm washing and swim-up was found to significantly improve sperm motility for men of infertile couples and the increment of percent sperm motility after sperm preparation allowed significant differentiation of pregnant and nonpregnant patients in asthenozoospermia(submotile) group (p<0.01). The author suggest that the increment of percent sperm motility after sperm washing and swim-up could be a useful screening tool for in vitro procedure proposed to improve fertility in the intrauterine insemination of asthenozoospermia.

• Korean Journal of Animal Reproduction
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• v.15 no.3
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• pp.201-205
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• 1991

This study was carried out to investigate the effects of Ca and BSA on hydrogen ion concentration in sperm washed solution. The results obtained were as follows : 1. The hydrogen concentration in 1st and 2nd sperm washed solutions was signifcinatly(p<0.01) higher when sperm was washed with SHPsolution containing 2mM Ca than when sperm washed with SHP solution or SHP solution containing 10mM Ca. 2. The hydrogen ion concentration in sperm washed solution was significnatly(p<0.05) higher when seprm was washed with SHP solution containing BSA-FAF than when sperm was washed with SHP solution or SHP solution containing BSA-V.

• Asian-Australasian Journal of Animal Sciences
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• v.21 no.8
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• pp.1127-1133
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• 2008

This study was designed to develop a method of liquid storage of boar sperm at $4^{\circ}C$ by using the modified Beltsville F5 (BF5) diluent with bovine serum albumin (BSA) and N-acetyl-D-glucosamine. Boar sperm were stored in lactose-egg yolk and N-acetyl-D-glucosamine (LEN), BF5 and Golden-Pig liquid 4 (GPL4) diluents at $4^{\circ}C$ for 5 days and were examined for sperm viability, adenosine triphosphate (ATP) and in vitro fertilization (IVF). The percentage of sperm viability in GPL4 diluent was higher than in LEN and BF5 diluent from 1 to 5 days of storage at $4^{\circ}C$. The percentage of sperm viability steadily declined from 1 to 5 days of storage in the three different diluents. Sperm ATP in GPL4 diluent was higher than in LEN and BF5 diluents from 1 to 5 days of storage. Sperm ATP rapidly declined after 5 days of storage in the three different diluents. Porcine oocytes matured in vitro were inseminated with different sperm concentrations of liquid semen stored for 3 days in GPL4 diluent. The percentage of monospermic oocytes did not show any differences from 2.5 to $20{\times}10^5$ sperm/ml. However, the percentage of polyspermic oocytes in the sperm concentration of $2.5{\times}10^5$ sperm/ml was lower than in concentrations of 5, 10 and $20{\times}10^5$5 sperm/ml. The percentage of blastocysts from the cleaved oocytes at $2.5{\times}10^5/ml$ sperm concentration was significantly lower than at 5, 10 and $20{\times}10^5sperm/ml$ concentrations. In conclusion, GPL4 diluent can be stored at $4^{\circ}C$ for 5 days and showed higher sperm viability and sperm ATP concentration compared with LEN and BF5 diluents. Also, we found that GPL4 diluent can be used for IVF of porcine oocytes.

• Journal of Embryo Transfer
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• v.31 no.2
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• pp.109-116
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• 2016

The recovery of epididymal sperm in animals is considered as one of the important tools to preserve high value or endangered species. However, there are no appropriate castrating indicators such as months of age in bull, sperm morphology, and motility, particularly in young Korean native bull (Hanwoo). Therefore, this study aimed to investigate sperm number, morphology, and motility of sperm in the epididymis tail of young Hanwoo bulls at 8 and 15 months of age. After castration, epididymal tails were collected and minced with blades to recover sperm. In experiments 1 and 2, sperm number, morphology, and motility were examined. Total number of sperm and percentage of normal sperm from bulls at 8 months of age was lower than that of bulls at 15 months of age after collection (P<0.05). Percentage of abnormal head, tail, proximal cytoplasmic droplet, dead and damaged acrosome of sperm from bulls at 8 months of age were higher than those of bulls at 15 months of age (P<0.05). In experiment 3, sperm motility from bulls at 8 and 15 months of age were examined before freezing and after thawing. Frozen-thawed sperm at 8 months of age showed low total motility and motile sperm with ${\geq}25{\mu}m/sec$ compared to those at 15 months of age and commercially-used sperm (P<0.05). In conclusion, sperm derived from the epididymal tail of bulls at 8 months of age showed high abnormal morphology and poor motility, which are not adequate for AI and IVF. On the other hand, sperm derived from the epididymal tail of bulls at 15 months of age showed high normal morphology and motility.

• Clinical and Experimental Reproductive Medicine
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• v.27 no.3
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• pp.267-273
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• 2000

Objectives: We have previous reported that thawed testicular sperm and sperm extracted from seminiferous tubule could achieved optimal fertilization and pregnancy in azoospermic patients. However, thawed testicular sperm did not show motility in many cases. Therefore we studied viability of immotile sperm extracted from frozen-thawed seminiferous tubule using hypo-osmotic swelling (HOS) test and eosin-Y test. Materials and Methods: After sperm extraction using for ICSI, the remained sections of seminiferous tubules were frozen with a computerized freezer. For thawing and preparation of testicular sperm, the seminiferous tubules were thawed by removing from $LN_2$ and letting them at room temperature for 10 min followed by %37^{\circ}C$ water bath for 10 min. The prepared samples were washed for free of preservation medium and sperm preparation method described previous. Sperm was suspended in 0.1 ml hypoosmotic solution. After 30 minutes, the type of distally coiled sperm were assessed. Results: In 44 cases of cryopreservation of seminiferous tubules in obstructive azoospennic patients, the fertilization rates with 2PN were 71.4% and pregnancy rates were 34.1%. The presence of motile spermatozoa on subsequent post-thaw testicular sperm remarked 15.1% and were increased to 77.3% just before ICSI. After sperm extracted from frozen-thawed seminiferous tubule, 3 hrs later in in vitro culture, the cases of presence of motile sperm, reaction of hypo-osmotic swelling test and viable sperm were 63.6% (28/44), 93.2% (41/44), and 77.3% (34/44), respectively. Conclusions: Just after post-thawed testicular sperm did not showed motility. Although motility was gained after in vitro culture, many cases showed non-motile sperm until optimal insemination time. However, HOS test showed positive reaction in non-motile sperm. Therefore, HOS test is an alternative method for the selection of viable sperm for ICSI.

• Korean Journal of Animal Reproduction
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• v.10 no.2
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• pp.192-203
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• 1986

Two experiments in this study were designed to compare the potential for in vitro capacitation and in vitro fertilization of ejaculated sperm among individual rabbit bucks. In experiment 1, for in vitro capacitation, the ejaculated sperm were preincubated in DM for 12 hr or 18 hr after HIS treatment, then 12 hr -or 18 hr- preincubated sperm were incubated with superovulated rabbit ova in a 5% CO2 incubator for 36 hr at 38$^{\circ}C$, and a part of cleaved ova was transferred to the recipient does for implantation of embryo. In experiment 2, effect of lysolecithin addition to preincubation medium on induction of accelerated in vitro capacitation and in vitro fertilization of individual rabbit sperm was studied. Experiment 1; 1. Percent acrosome reaction of sperm, noted after staining, after 12 hr or 18 hr preincubation ranged from 52.5 to 76.0% and from 67.5 to 90.0%, respectively and sperm motility index of these sperm ranged from 20.0 to 47.5 for 12 hr-preincubated sperm and from 15.0 to 37.5 for 18 hr- preincubated sperm. There was no a certain relation between percent acrosome reaction and sperm motility index. 2. In vitro fertilization rate (cleavage rate) of in vitro capacitated sperm varied widely among individual bucks, ranging from 0 to 47.8% for 12 hr - preincubated sperm and from 0 to 60.9% for 18 hr -prein- cubated sperm. Cleavage rate of 18 hr - preincubated sperm was higher and faster than that of 12 hr - preincubated sperm. 3. Eight of 44 in vitro fertilized embryos transferred into 6 recipients were implanted in 4 recipients (66.7%) up to day 15 and implnatation rate was 18.2%. Experiment 2; 1. The percent acrosome reaction of sperm before and after 4 hr preincubation in DM without lysolecithin varied significantly among individual bucks, ranging from 0.4 to 18.4% and from 1.7 to 37.4%, respectively and percent acrosome reaction of sperm at 30 min after addition of 60${\mu}$g/ml lysolecithin also was significantly different among bucks, ranging from 19.2 to 67.1%. 2. Effect of accelerated acrosome reaction following lysolecithin addition was more considerable in the individuals showed less percent acrosome reaction before and after 4 hr preincubation. Percentage of motile sperm and motility score showed a trendency towards a decrease with increase of preincubation time and time after lysolecithin addition. 3. In vitro fertilization rate (cleavage rate) at 24 hr postinesmination with pooled sperm were treated to 60 $\mu\textrm{g}$/ml lysolecithin for 30 min after 4 hr preincubation was 24.6%, a higher rate than 13.2% for control. While 80 $\mu\textrm{g}$/ml lysolecithin-added sperm showed a lower cleavage than control and 60$\mu\textrm{g}$/ml-added sperm at both 24 hr and 48 hr postinsemination. These results from 2 experiments suggest that more useful preincubation time for the in vitro capacitation of ejaculated rabbit sperm is 18 hr in DM after HIS treatment, although there is wide variation in vitro capacitation and in vitro fertilization rate among individual bucks, and lysolecithin addition to at least 4 hr - preincubated sperm in DM can result in almost same in vitro fertilization rate as that of 18 hr - preincubated sperm in the experiment 1.

• Asian-Australasian Journal of Animal Sciences
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• v.13 no.1
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• pp.10-18
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• 2000

This paper describes the magnetic orientation of the intact and demembranated bull sperm treated by DTT or heparin in a 5,400 G static field. Semen samples collected from four bulls (Japanese Black) were mixed to the same sperm density. One percentage triton X-100 was used to extract the plasma membrane. The intact and demembranated sperm suspensions were treated with 20, 200, 2,000 mM DTT, 100, 1,000 or 10,000 units heparin solutions at $4{^{\circ}C}$ for 6 days. The decondensation of the sperm nuclei treated by DTT or heparin was examined by measuring the sperm head area at 1, 3, and 6 days. After measuring the area, each sperm sample was exposed to a 5,400 G static magnetic field generated by Nd-Fe-B permanent magnets for 24 hours at room temperature. Results showed that the decondensation of bull sperm nuclei was not induced by the heparin treatment, however, incomplete decondensation was induced by the DTT treatment. During the magnetic orientation, bull sperms treated by DTT or heparin had low percentages of long axis perpendicular to the magnetic lines of force. However, different aspects were obtained for long axis perpendicular orientations following treatment of DTT or heparin. Through the DTT treatment, the decline of long axis perpendicularly oriented percentages was due to the increase of long axis parallel orientation with the head of the flat plane perpendicular to the magnetic lines of force, whereas, using the heparin treatment, the decline of long axis perpendicular orientation was due to the increment of long axis parallel orientation with the head of the flat plane parallel to the magnetic lines of force. Also, percentages of the head of the flat plane perpendicular were decreased by the heparin treatment. These findings suggest that maintaining the structure of protamine in the chromatin is necessary for the sperm head to orient with its flat plane perpendicular, and maintaining the disulfide bond in the chromatin is necessary for the long axis of sperm to orient perpendicularly.