• Clinical and Experimental Reproductive Medicine
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• v.16 no.1
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• pp.15-21
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• 1989

An in vitro fertilization assay employing zona-free hamster embryos was used to investigate human spermatozoal fertilitzing ability. Yanaghimarchi et al.(1976) first introduced this cross species fertilzation technique, with its application as a diagnostic tool for male infertility. Human spermatozoa were preincubated for 3 to 4 hrs in B W W medium at concentration of $4{\times}10^6$ sperm/ml prior to the addition to zona-free hamster embryos. After 3 hrs, human sperm was evaluated for fertilizing potential by the presence of swelling or decondencing sperm head in the cytoplasm. The results of penetration rates for sperm were as follow : 1. The average penetration rate of a 7 fertile donor group was $47.8{\pm}27.67%$(Range 14.3-98.0%) 2. The average penetration rate of 12 infertile patients with normal semen analysis was $21.7{\pm}26.9%$(Range 0-38.8%) 3. The average penetration rate of 10 infertile patients with semen abnormalities was $6.1{\pm}8.1%$(Range 0-25%)

• Korean Journal of Animal Reproduction
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• v.9 no.2
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• pp.148-152
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• 1985

When goat spermatozoa were preincubated for 4-6 h and 6 h in the uteri isolated from hamster and rat, and for 6 h in the hamster uterus in situ, they developed the ability to penetrate zona-free hamster eggs in vitro. Zona-free hamster eggs were not penetrated after insemination with goat spermatozoa preincubated in the isolated hamster uterus 4 h before and 2 h after expected time of ovulation, respectively. Zona-free hamster eggs were not penetrated after insemination with goat spermatozoa preincubated for 4 h in the isolated hamster uterus, but 10 and 18% of eggs were penetrated by spermatozoa preincubated for 5 and 6 h in the isolated uterus.

• Korean Journal of Animal Reproduction
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• v.9 no.2
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• pp.153-157
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• 1985

Ejaculated and epidiymal goat spermatozoa were preserved for 0, 6, 12 adn 18 h, and 0 and 18 h in a semi-aerobic condition at 20-$25^{\circ}C$, and preincubated for 5-6 h in a CO2 incubator in m-KRB solution. Then they were preincubated at different concentrations (3-5, 25-48 and 105-190$\times$107/ml), and ability of penetration into zona-free hamster eggs in vitro was examined. When ejaculated spermatozoa were preincubated in m-KRB solution after presservation for 12 and 18 h, 12 and 29% of zona-free eggs were penetrated, and only 4% of eggs were penetrated by epididymal spermatozoa which were preincubated after preservation for 18 h. When spermatozoa were preincubated at a low concentration, the penetration rates were very low. But when the sperm concentration during preincubation was 25-48 and 105-190$\times$107/ml, the penetration rates increased to about 30%.

• Reproductive and Developmental Biology
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• v.36 no.3
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• pp.155-161
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• 2012

Germ cell-specific hyaluronidases such as sperm adhesion molecule 1 (SPAM1) and hyaluronoglucosaminidase 5 (Hyal5) are in part responsible for dispersal of the cumulus cell mass, which is a critical step in establishing fertilization in mammals. In this study, we identified two testis-hyaluronidases, SPAM1 and Hyal5, in hamster and rat. These two genes were expressed specifically in the testis. At the protein level, hamster SPAM1 and Hyal5 display 78.7% and 75.4% identity with mouse SPAM1 and Hyal5. Further, the activity of the enzymes with respect to cumulus cell dispersion did not differ, although we observed that the enzymatic activity differed in pH range. These studies suggest that different sperm hyaluronidases are capable of dispersing the cumulus cell mass despite differences in enzyme activity.

• Korean Journal of Animal Reproduction
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• v.10 no.1
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• pp.91-99
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• 1986

This experiment was undertaken to examine the effects of HIS treatment on the motility and acrosome reaction of frozen bovine spermatozoa and to test their abilities to interact with zona-free hamster eggs in vitro. Also, in vitro results were compared with those of bull's fertility in AI. The frozen semen from four Holstein bulls were exposed to HIS-DM for 5 minutes after thawing and then preincubated for 60 minutes in DM prior to insemination. The hamster eggs were mounted, fixed and stained 6 hours after exposure to boving spermatozoa and examined under a phase-contrast microscope. 1. The sperm motility expressed as a mobility index dro, pp.d significantly from 60-75 to 12-24 after exposure to HIS-DM, but increased in 32 to 41 at insemination. Bull C showed a low motility index than those of the orher bulls. The percentage of acrosome reaction by staining procedure were increased by HIS-DM treatment but did not change during 7 hours incubation period in DM. 2. The overall percentage of hamster eggs interacting with bull spermatozoa was 56.3%, 58.3%, 66.6% and 70.0%, respectively. Although there was no significant difference among bulls in the penetration rate of spermatozoa into hamster eggs, high proportions of eggs interacted with spermatozoa from Bull C and D than those from Bull A and B. 3. The conception rates (60-90 day RP) resulting from AI were 62.5%, 67.5% and 70.9% for Bull A, B and C, respectively. These results were in good agreement with the invitro results that the proportions of bull sperm-egg interction were greater for Bull C than for Bull A and B.

• Korean Journal of Veterinary Research
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• v.39 no.1
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• pp.189-203
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• 1999

To determine sex of bovine embryos using hamster histocompatibility Y(H-Y) antibodies, bovine compact morulae were incubated for 6 hours in TCM199 supplemented with 10% hamster H-Y antiserum and the embryos with developmental arrest were diagnosed as male embryos, while the embryos showing development during the incubation as female embryos. This presumptive embryo sexing was confirmed by polymerase chain reaction(PCR)method. 1. In the result of hamster sperm cytotoxicity test to measure H-Y antibody titer, the rate of dead sperm was considerably lower in H-Y antiserum absorbed with hamster male splenocytes than in H-Y antiserum absorbed with hamster female splenocytes or H-Y antiserum unabsorbed with splenocytes(p<0.01). 2. The rate of oocytes fertilized in vitro and the rate of blastocysts of the fertilized oocytes were 58.5% and 32.4%, respectively. The rate of blastocysts on day 8 was 15.9%, denoting the highest rate during whole culture period posterior to in vitro fertilization (IVF). 3. The bovine 16 cell and compact morulae embryos incubated in the medium supplemented with hamster H-Y antibodies showed 37.1% and 48.9% of developmental arrest which were diagnosed as male, respectively, and rates of redeveloped embryos from the arrested were 24.1% in 16 cell and 44.3% in compact morulae embryos, respectively, denoting higher rate of sex determination and rate of redevelopment in compact morulae than 16 cell embryos. 4. Bovine compact morulae of Korean cattle and Holstein were treated with hamster H-Y antibodies for sex determination and the rates of developmental arrest(diagnosed as male) were 48.4% for Korean cattle and 47.9% for Holstein, respectively. The rates of redeveloped embryos to blastocyst after treatment were 42.6% for Korean cattle and 41.8% for Holstein, respectively, showing no significant differences of sex determination and redevelopment between both breed. 5. The sex determination of bovine embryos(Korean cattle and Holstein) using hamster H-Y antibodies was diagnosed by PCR for confirmation, denoting the rates of 86.1% for Korean cattle and 85.9% for Holstein male embryos, respectively, and the rates of 91.9% for Korean cattle and 90.1% for Holstein female embryos, respectively, with no significant differences of sex determination between both breed. These results indicated that hamster H-Y antibodies can be usable for sex determination of bovine embryos of Korean cattle and Holstein, the viability of bovine embryos was sustained while being cultured in the medium supplemented with hamster H-Y antibodies of appropriate titer and sex determination of bovine embryos by PCR can be feasible for confirmation.

• Korean Journal of Animal Reproduction
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• v.23 no.1
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• pp.29-36
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• 1999

This study were performed the effect of methylxanthine derivatives on hamster epididymal sperm motility. To assess the effect of methylxanthine derivatives on motility kinematics of epididymal sperm, pentoxifylline, 2-deoxy-adenosine and hypoxanthine were added to TALP medium. 1 mM of pentoxifylline, significantly increased VCL, VAP, VSL of spermatozoa obtained from corpus and cauda epididymis. With 1 mM of 2-deoxyadenosine, VCL, VAP, VSL of spermatozoa obtained from corpus and cauda epididymis were significantly increased, but 2 mM ADE for cauda spermatozoa was effective than 1 mM. In the case of hypoxanthine, various concentrations were not significantly effective, but 2 mM HX showed higher effect than other concentrations. pentoxifylline 1 mM and 2-deoxyadenosine 1 mM significantly increased VCL, VAP of corpus and cauda epididymal spermatozoa and VSL of cauda epididymal spermatozoa. From these results it could be concluded that the addition of methylxanthine increase motility kinematics of hamster spermatozoa collected from epididymis.

• Clinical and Experimental Reproductive Medicine
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• v.18 no.1
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• pp.63-71
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• 1991

To establish the normal fertile range in the results of the sperm zona-free hamster ova penetration assay (SPA) in Korean male, SPA using the low temperature ($4^{\circ}C$) capacitation in TEST-yolk buffer (TYB) was performed in 67 fertile and 26 infertile men. Sperm parameters in routine semen analysis were also checked and compared with the results of SPA. Sperm concentration, motility and motility index (MI) were significantly higher in fertile group compared with infertile group: $96.0{\pm}46.6$ vs $43.6{\pm}31.9{\times}10^6/ml$, $65.5{\pm}14.8%$ vs $45.8{\pm}23.6%$ and $46.31{\pm}13.29$ vs 27.40{\pm}17.98$, respectively. In fertile group, the hamster ova penetration rate (PR) was $98.5{\pm}5.0%$ (80%-100%), and the penetration index (mean penetrations per ovum, PI) was $9.59{\pm}6.35$(3.1-29.0). All the fertile men showed PI>3.0. In infertile group, PR was $24.6{\pm}24.8%$ (0%-70%), and PI was $0.40{\pm}0.42$ (0-1.3). Both PR and PI were significantly lower in infertile group. There was a significant correlation beween PI and sperm motility or MI, respectively, in fertile group whereas there was no correlation in infertile group. These data suggest that SPA using the low temperature capacitation in TYB can be a valuable diagnostic tool for the assessment of male fertility in vitro and provide an important supplement to the traditional tests of sperm quality.

• Clinical and Experimental Reproductive Medicine
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• v.18 no.1
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• pp.73-80
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• 1991

We compared fertilizing potential measurements by the zona-free hamster egg penetration assay with the in vitro fertilization and embryo transfer program was evaevulated for their ability to fertilize zona free hamster egg. Spermatozoa from 12 presumeably fertile donors and from the male partners of 56 infertile couples were evaluated for their ability to fertilizing potentials. Penertration rates of fertile donors were $36.2{\pm}27.7%$ ; Fertilization rates of infertile couples between with normal semen parameters and with abnormal semen parameters were $28.7{\pm}19.1$, $5.7{\pm}8.9%$, respectively. Sperm motility of couples with penetration rates between on 15-30% and on 30> were $54.1{\pm}4.6$, $55.5{\pm}8.3%$ respectively. Hamster penetration rates of couples participating in an in vitro fertilization and embryo transfer program was $38.9{\pm}29.9%$. But in one case, a positive fertility assessment was obtained in the absence of fertilization of the wife's eggs attributable to egg immaturity. This method may have potential value as a diagnostic tool in evaluation human sperm fertilization capacity which avoids the ethical and logistical problems associated with fertilizing of human eggs in vitro.

• Clinical and Experimental Reproductive Medicine
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• v.29 no.1
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• pp.57-66
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• 2002

Objective : This study was designed to investigate the interrelationship and clinical usefulness of sperm morphology by strict criteria (SM), acrosome reaction following ionophore challenge test (ARIC) and sperm penetration assay (SPA) using zona-free hamster ova as prognostic factors in in vitro fertilization. Materials and Methods: Semen samples were provided by 83 patients undergoing IVF. We first evaluated the differences between normal fertilization group and poor fertilization group on three andrologic tests. Secondly, we analyzed the relationship between the three andrologic tests and in vitro fertilization on IVF settings. Finally, we evaluated the effectiveness of the three andrologic tests as the prognostic indicators for fertilizing ability. Results: The fertilization rate of all men in the poor fertilization group was less than 30%; but there was no evidence that this poor fertilization was due to oocyte defects. The results of three andrologic tests were significatly higher in normal fertilization group. Fertilization rate (%) in vitro was highly correlated (p<0.001) with % normal sperm by SM, ARIC value (%), and SPA result. By using Receiver-Operator-Characteristic curve (ROC), we evaluated the effectiveness of these three tests. The sensitivity and specificity of SM, ARIC test and SPA in predicting fertilization potential in IVF setting were 76% and 75%, 84% and 90%, and 76% and 95%, respectively. Conclusion: Our data suggest that the three andrologic tests can be reliable tools as prognostic factors of sperm fertilizing ability. Among these test, ARIC test and SPA gave more accurate information on fertilizing capacity. ARIC test was shown to have a predictive value for fertilizing ability comparable to that of SPA that appears to be a simple and cost-effective addition to current andrology laboratory. Combined application of these three tests may give more information on predicting sperm fertilizing capacity.