• 한국수정란이식학회지
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• 제19권3호
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• pp.265-273
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• 2004

본 연구는 mNCSU-23에서 체외성숙시킨 난자를 이용하여 돼지 체외수정시 적절한 배양액과 정자농도를 구명하고자 수행하였다. 정액은 액상정액을 이용하였고 체외수정배양액으로 mTBM과 mTLP-PVA, 정자농도는 운동정자수 5${\times}$$10^4$, 1${\times}$$10^{5}$ , 5${\times}$$10^{5}$ sperm/$m\ell$로 체외수정 한 다음 NCSU-23에서 체외발달을 유도한 결과는 다음과 같다. 1. 돼지 체외성숙란을 mTBM 또는 mTLP-PVA에서 운동정자수 1${\times}$$10^{5}$ sperm/$m\ell$로 체외수정시킨 다음 체외발달시킨 결과, 정자침투율, 웅성전핵형성율, 다정자수정발생율, 난분할율, 배반포 발달율 등 어느 것도 두 배양액간 차이를 나타내지 않았다. 2. 체외성숙란을 mTBM에서 운동정자수 5${\times}$$10^4$, 1${\times}$$10^{5}$ , 5${\times}$$10^{5}$ sperm/$m\ell$l로 체외수정시킨 다음 체외발달시킨 결과, 정자침투율, 웅성전 핵형성율, 다정자수정발생율 등은 모두 정자 농도간 유의적인 차이를 보였고(P<0.05), 난 분할율과 배 반포발달율은 5${\times}$$10^4$이 1${\times}$$10^{5}$ , 5${\times}$$10^{5}$ sperm/$m\ell$보다 유의 적으로 낮았으나(P<0.01), 1${\times}$$10^{5}$ 와 5${\times}$$10^{5}$ sperm/$m\ell$ 간에는 차이가 없었다. 따라서, 액상정액을 이용한 돼지 체외성숙란의 체외수정배양액으로 mTBM과 mTLP-PVA간 차이는 없으며, mTBM에서 체외수정시킬 때 정자농도는 운동정자수로 1${\times}$$10^{5}$ sperm/$m\ell$가 적절할 것으로 사료된다.

• 한국환경보건학회:학술대회논문집
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• 한국환경보건학회 2005년도 국제학술대회
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• pp.319-322
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• 2005

Porous building materials are not only sources of indoor air pollutants such as volatile organic compounds (VOCs) but they are also strong sinks of these pollutants. Volatile organic compounds have been implicated in impaired spermatogenesis, increase in the incidence of malformed sperm and decrease in the percentage of moving sperm. The aim of this study was to determine and compare the direct effects of various volatile organic compounds (phenol, formaldehyde; HCHO, ethanol, toluene, styrene) on motility and survival rate of human sperm in vitro. Semen samples from 3 health subjects were prepared using swim-up method and 1-10mM volatile organic compounds were added to the test medium. HCHO and phenol produced significant decreases in the motility and survival rate with a different potency. The most potent inhibition of motility and survival rate was observed after exposure to HCHO. Less than 1mM HCHO significantly inhibited sperm motility. When ethanol is added directly to sperm, at concentrations equivalent to that in serum after heavy drinking, these damaging effects were lowest compared with other volatile organic compounds. Present study shows that each compound has differential toxic potency to human sperm and we need special caution for the use of HCHO and phenol.

• 한국수정란이식학회지
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• 제25권4호
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• pp.237-245
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• 2010

Only an optimum number of viable spermatozoa in a frozen-thawed insemination dose can ensure conception at artificial insemination (AI). We report here the percentages of normal, abnormal and viable spermatozoa present in the frozen-thawed semen of 20 Black Bengal bucks used for commercial AI. Bucks in this experiment were of 19.3~46.1 months old and 25~42 kg body weight. Four semen straws (0.25 ml) from each buck were collected for evaluation of their kinetic parameters. Scrotal circumference was measured by using a scrotal tape, sperm motility was estimated on eye estimation and sperm concentration was determined by using a haemocytometer. Sperm morphology was studied in paraformaldehyde fixed spermatozoa under differential interference contrast (DIC) microscope. To determine the proportion of live (plasma membrane intact) spermatozoa, semen was stained with SYBR-14 and propidium iodide and examined under fluorescent microscope. Scrotal circumference, post-thaw sperm motility, sperm concentration per insemination dose and proportion of normal spermatozoa were $21.5{\pm}0.7\;cm$, $43.5 {\pm}5.4%$, $83.5{\pm}6.7$ million and $88.3{\pm}4.1%$, respectively. The percentages of spermatozoa with head shape and acrosome abnormalities were lower ($2.7{\pm}1.1$ and $1.4{\pm}1.3$, respectively), whereas higher percentages of abnormalities ($7.0{\pm}1.8$) were observed in mid piece and tail portion. The proportion of live spermatozoa was $28.5{\pm}5.4$. It is concluded that although a good number of morphologically normal spermatozoa are present in the insemination dose, the proportion of live spermatozoa is low, which warrants further improvements of buck semen freezing procedures to ensure good quality at AI.

• 한국수정란이식학회지
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• 제27권1호
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• pp.37-43
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• 2012

This study evaluated a method of sorting X and Y chromosomes based on size using the forward angle light scatter related refractive index (FSC) of a flow cytometer. Hanwoo bulls sperm were separated to X and Y chromosomes by the parameters of FSC or Hoechst 33342 intensity. As a result, using monitor program linked flow cytometry during sorting processing, the purities were $97{\pm}0.57$ or $96{\pm}0.67%$ for the X-fraction and $96{\pm}0.33$ or $97{\pm}1.33%$ for the Y-fraction in the two sperm sorting methods. There were no differences in the X and Y ratios (X and Y %) between the sperm sorting methods based on FSC or DNA content. The proportions of female and male embryos used for in vitro fertilization and development were $66.03{\pm}3.31$ or $69.37{\pm}1.41%$, and $70.56{\pm}2.42$ or $56.11{\pm}3.09%$ when sperm were processed using the sex sorting method by FSC or Hoechst 33342. In conclusion, further study is needed to determine the optimum procedure and improve the nozzle to enhancing sorting accuracy or efficiency. Also, the findings of this study do not negate the possibility that the difference method of sperm sorting cannot use a UV laser beam.

• 대한한방부인과학회지
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• 제19권1호
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• pp.111-124
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• 2006

Purpose : This study was conducted to investigate the dose dependent effects of Panax Ginseng on the reproductive functions in mice. Methods : We used the 8-week-old mice, and administered 0.2ml extract solution of Panax Ginseng in the different concentration once a day for 60 days. The control group was administered 0.2ml normal saline in the same way and duration. We counted the total, motile and normal sperm number of the cauda epididymis and measured the activities of sperm hyaluronidase, peroxidase and catalase of the isolated testis tissues. And we observed histological changes of surgically isolated testis by histochemical methods. Results : All Panax Ginseng extract solution groups showed significantly dose dependent differences in the total number, the motility and normality of sperms compared with the control group, respectively. In the histological analysis of the testicular tissues, all Panax Ginseng extract solution groups showed the enlargement of testicular lobe diameter and apparent angiogenesis between seminiferous tubules. And the activity of typical sperm enzyme, hyaluronidase, was significantly increased in the Panax Ginseng extract solution groups compared to the control group. In the antioxidant activity analysis, the activities of peroxidase and catalase were significantly increased in the Panax Ginseng extract solution groups compared to the control group. Conclusion : This study shows that Panax Giseng has the beneficial effects on the concentration, morphology and motility of sperm, the activities of sperm hyaluronidase, testicular peroxidase and catalase. We can suggest that Panax Ginseng be useful for the treatment of male infertility.

• Clinical and Experimental Reproductive Medicine
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• 제48권1호
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• pp.27-33
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• 2021

Objective: The chief outcome of testicular torsion in clinical and experimental contexts is testicular ischemia. Curcumin, a compound with anti-inflammatory and antioxidant properties, has fascinated researchers and clinicians for its promise in the treatment of fertility diseases. Methods: Thirty-five fully grown male mice were randomly classified into five groups: control, sham, testicular torsion, treatment group 1 (testicular torsion+short-term curcumin), and treatment group 2 (testicular torsion+long-term curcumin). Thirty-five days later, spermatozoa from the right cauda epididymis were analyzed with regard to count and motility. Toluidine blue (TB), aniline blue (AB), and chromomycin A3 (CMA3) staining assays were used to evaluate the sperm chromatin integrity. In addition, the terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick-end labeling (TUNEL) test was used to assess apoptosis. Results: Treatment group 1 exhibited a remarkably elevated sperm count compared to the testicular torsion group. Additionally, notably lower sperm motility was found in the testicular torsion group compared to the control, treatment 1, and treatment 2 groups. Staining (CMA3, AB, and TB) and the TUNEL test indicated significantly greater testicular torsion in the torsion group compared to the control group (p<0.05). The data also revealed notably lower results of all sperm chromatin assays and lower apoptosis in both treatment groups relative to the testicular torsion group (p<0.05). Significantly elevated (p<0.05) AB and TB results were noted in treatment group 1 compared to treatment group 2. Conclusion: Curcumin can compensate for the harmful effects of testicular ischemia and improve sperm chromatin quality in mice.

• Asian-Australasian Journal of Animal Sciences
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• 제23권1호
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• pp.33-40
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• 2010

In this study, the production of transgenic goats using sperm to integrate exogenous DNA and artificial insemination (AI) was carried out and the technical protocols for sperm-mediated gene transfer (SMGT) in the goat were optimized. The standard sperm parameters and the ability to bind foreign genes were assessed to select suitable sperm donor bucks. A total of 134 oestrous does were divided into 4 groups and inseminated using different methods and sperm numbers. The does of Groups I to III were inseminated with fresh semen ($1-2\times10^{7}$ and $10^{6}$ sperm) or frozen-thawed semen ($10^{6}$ sperm), respectively, through conventional intra-cervical AI, and the does of Group IV with frozen-thawed semen ($10^{6}$ sperm) through intrauterine AI. Total genomic DNAs were extracted from ear biopsies of the offspring. The presence of $pEGFP-N_{1}$ DNA was screened by PCR and then by Southern blotting analysis. A total of 76 live kids were produced and 8 kids were tested transgene positive on the basis of agarose gel electrophoresis of the PCR-amplified fragment. Southern blotting analysis of the samples showed 5 positive kids. A transgenic ratio of 10.53% was detected using PCR and 6.58% using Southern blotting. The positive kid rate assayed by PCR and Southern blotting of frozen-thawed goat semen was 3.61% and 9.27% higher than that of untreated semen. The results show that transgenic goats can be produced efficiently by the method of artificial insemination using sperm cells to integrate the exogenous DNA and intrauterine insemination allowed low numbers of DNA-transfected spermatozoa to be used, with satisfactory fertility.

• Clinical and Experimental Reproductive Medicine
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• 제23권3호
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• pp.351-355
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• 1996

Objective: To analyze the direct effect of nitre oxide, generated from sodium nitroprusside, on sperm motility and reactive oxygen species. Design: Human sperm samples were treated to allow swim-up and washing. And the samples were devided into four aliquots. Each aliquot was incubated with either concentration at 0, 100nM, $10{\mu}M$, 1mM of nitroprusside. Intervention: Samples were measured chemiluminosence for reactive oxygen species of each aliquot with concentrations at 0, 100nM, $10{\mu}M$, 1mM of nitroprusside at allowing swim-up and washing of sperm. Main Outcome Measures: Percent motion parameters and viability were asse-ssed at 0, 3, 6, 12, 24 hours incubation. Results: The percent viablity was lower slightly in control group (50.2%) than that in sperm treated with 100nM of nitroprusside(57.5%) at 24 hours after incubation, while was reduced significantly in sperm with concentra-tion of $10{\mu}M(42.1%)$ and 1mM(21.3%)of nitroprusside at 6 hours after incubation. And the sperm treated with 1mM of nitroprusside was immotile totally at 6 hours after incubation. The straight line$(35.3{\pm}5.6%)$, the rapid forward$(37.2{\pm}6.4%)$ and the weak curvilinear velocity$(9.6{\pm}2.4%)$were more favorable comparing with those ($32.4{\pm}4.2%$, $30.0{\pm}7.8%$ and $18.0{\pm}4.6%$ respectively) in control group at 3 hours after incubation, but reduced significantly in sperm treated with $10{\mu}M$ and 1mM of nitroprusside. The levels of reactive oxygen species in control(700 c.p.m.) is lower significantly than that in each experimental groups of sperm treated with nitroprusside. And the levels of reactive oxygen species were 2200 c.p.m. in 100nM, 6200c.p.m. in $1{\mu}M$ and 12800c.p.m. in 1mM respectively. Conclusion: These results suggested that the concentration of 100nM of nitroprusside on sperm is beneficial to the maintanance of viablity and motile velocity, but detriment in high concentration of $10{\mu}M$ or 1mM of nitroprusside.

• Reproductive and Developmental Biology
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• 제31권3호
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• pp.139-143
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• 2007

본 연구는 체세포를 이용하여 생산된 복제 한우 수소의 번식능력을 검토하기 위해 실시하였다. 복제 한우 수소(C-38 및 C-39) 또는 일반 한우 종모우로부터 정액을 채취하여 정자의 수 및 동결 전 후의 생존성 등을 살펴보았으며, 정자의 운동성 등은 computer assisted sperm analysis(CASA)를 이용하여 측정하였다. 또한, 이들의 수정 능력을 확인하기 위하여 체외수정과 인공수정을 각각 실시하였다. 정액 성상에서는 복제 수소들과 일반 종모우 간에 정액의 양, 정자의 농도 및 동결융해 후의 생존성 등에서 차이가 나타나지 않았다. CASA를 이용한 분석에서 운동성, 곡선 운동 속도(VCL), 직선운동 속도(VSL) 및 평균 진행 속도(VAP) 등은 복제 수소의 정액이 일반 종모우의 정액에 비하여 유의적으로 높았다 (p<0.05).체외수정에 따른 수정란의 분화율 및 배반포로의 발달율은 복제 수소와 일반종모우 간에 차이가 나타나지 않았다. 복제소 정액(C-38)을 이용하여 인공수정을 한 5두의 체세포 복제 대리모에서 암 수 각각 한 두씩의 건강한 복제 후대 송아지 2두를 생산하였다. 이상의 결과를 종합하여 보면, 실험에 공시된 복제 수소 개체간의 차이가 나타나기는 하였지만, 복제 수소는 정액 성상과 정자의 운동성 등에서 일반 종모우와 차이가 없었으며, 또한 인공수정을 통해 송아지를 생산함으로써 정상적인 번식능력이 있음을 확인하였다.

• Clinical and Experimental Reproductive Medicine
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• 제23권1호
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• pp.7-9
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• 1996

Here, A forty year old patient is presented, showing azoospermia following vasovasostomy. The bilateral testicular biopsies were performed to confirm whether ductal obstruction due to vasovasostomy or testicular failure existed. The finding of biopsy was spermatogenic arrest. After completion of medical testicular stimulation with clomiphene citrate and pentoxifilline for 3 months, repeat vasovasostomy was performed. Semenanaylsis revealed normal sperm parameters after operation. Necessity of testicular biopsy before deciding repeat vasovasostomy, accuracy of testicular biopsy, efficacy of testicular stimulation in the patient with spermatogenic arrest and effect of testicular biopsy on testicular fuction are discussed.