• 한국가축번식학회지
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• 제14권1호
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• pp.31-35
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• 1990

Eosin B의 사용에 의한 rat 난모세포의 생존성 및 발달가능성을 검사하였다. 염료는 Brinster's Medium for Ovum Culture puls glucose (BMOC)-2 medium ml당 eosin B 1.0mg을 함유하는 stock solution으로 준비하였다. 난모세포연구를 위하여 BMOC로 stock solution을 희석하여 만든 3가지의 염료농도들이 선택되었다 : 0.12mM, 0.60mM, 1.20mM. 난소세포들로부터 난모세포들을 채취하고, 보존기간 및 열 처리의 다양한 조건하에서 eosin B를 사용한 dye exclusion assay로 검사하였다. 생존한 난모세포는 eosin B를 배척하고, 사멸한 세포는 eosin B를 배척하지 않는 것으로 제시되었다.

• 한국수정란이식학회지
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• 제15권3호
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• pp.279-286
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• 2000

This study was conducted to develop an improved method for oocyte pick-up(OPU) with finger-sensibility using ultrasound-guidance from ovarian follicles in Holstein heifers. Oocytes were aspirated from ovarian follicles of clear-outline (>2mm), obscure-outline and invisible($\leq$ 2mm) on ultrasound images with 3 different vacuum pressure(40, 80, 120mmHg). Total number of oocytes recovered/follicles were 309/237(130.4%). 113/80(141.3%) and 107/74(144.6%) with 40, 80 and 120 mmHg of vacuum pressure, respectively. Mean number of oocytes recovered was higher in 2 OPU/week (18.3$\pm$5.3) than 1 OPU/week(14.5$\pm$4.1), but this difference was not statistical1y significant. The recovery rates were not affected by the number of OPU as 135.6%(282 oocytes/208 follicles) in 1~20 OPU, 137.7% (168/122) in 21~40 OPU and 148.4%(92/62) in 41~60 OPU, respectively. The proportions of good oocytes (Grades I) recovered were not significantly different by the number of OPU until 40 OPU(12.4% in 1~20 OPU vs 16.7% in 21~40 OPU). However, a significantly(P<0.05) lower recovery rate resulted from more than 40 OPU compared to less than 40 OPU(7.6%). These results imply that more fertilizable oocytes can be produced from invisible-immature follicles by transvaginal aspiration with finger-sensibility from Holstein heifers.

• 한국어류학회지
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• 제7권1호
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• pp.71-78
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• 1995

낙동강 상류의 여러 수계에서 거의 암컷으로 구성된 Cobitis sinensis - longicorpus complex 2배체와 3배체 집단이 높은 빈도로 출현하였다. 그 가운데, 수컷의 C. sinensis (2n=48) 및 C. longicorpus (2n=50)와 암컷 3배체 C. sinensis - longicorpus (3n=73)를 실험실내에서 인공 교잡 실험을 하였다. 그 결과 각 조합에서 얻은 자손의 체측 반문에는 부계 형질이 발현되었고 그들의 염색체는 모두 부계의 haploid genome이 포함된 diploid form (2n=48,49)이었다. 이상의 결과로 볼때 3배체 암컷인 Cobitis sinensis - longicorpus complex는 제 1 감수분열에서 uneven genome을 배제하고 제 2감수분열에 의한 반수체 ovum을 형성하는 독특한 생식 양상을 갖는 것으로 사료된다.

• 한국수정란이식학회지
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• 제29권1호
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• pp.35-41
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• 2014

Laparoscopic ovum pick-up (LOPU) is a convenient method for collecting oocytes in small ruminants. LOPU has the advantage of being a less invasive means of oocyte collection, thereby allowing for a repeated usage of the oocyte donor animals. A total of 25 Korean black goats were used in the winter season (December to February) and LOPU was applied to the goats which had been treated for superovulation more than two times during the last twelve months. Estrus was synchronized with an intravaginal insert containing 0.3 g progesterone for 10 to 12 days. Ovaries were hyperstimulated with eCG 1,000 IU oneshot, FSH with eCG (50 mg / 1,000 IU; 70 mg / 500 IU; 70 mg / 1,000 IU) oneshot or FSH multiple-shot with eCG oneshot ($20mg{\times}6/300IU$) given intramuscularly 72 h prior to LOPU. For these groups, the number of follicles (mean ${\pm}$ SEM) observed which developed to larger than 2 mm in diameter were $1.6{\pm}2.5$, $4.3{\pm}3.1$, $5.5{\pm}4.2$, $6.6{\pm}2.1$ and $8.8{\pm}7.8$, respectively. Oocytes were aspirated by using OPU needles and a vacuum pump. The overall oocyte retrieval rates were 41.4%. Oocytes were matured in TCM-199 supplemented with 10% (w/v) bovine serum albumin + $10{\mu}g/ml$ FSH + $1{\mu}g/ml$ $17{\beta}$-estradiol for 27 h at $39^{\circ}C$ in 5% $CO_2$ in air. Oocytes were parthenogenetically activated by ionomycin combined with 6-diethylaminopurine (6-DMAP). Total oocyte maturation and cleavage rate were 67.3% and 78.8%, respectively. In summary, LOPU is a useful oocyte collection method in Korean black goats that can provide immature oocytes for transgenesis or nuclear transfer.

• 생명과학회지
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• 제28권7호
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• pp.811-818
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• 2018

본 연구에서는 암컷 생식기관 내 축적된 수은 화합물의 위치와 시간에 따른 수은의 농도 변화를 확인하기 위하여 수행하였다. Methylmercuric chloride를 일주일에 한 번씩 총 3주간 사춘기 암컷 마우스에 피하 주사하였다. 시간에 따른 수은의 농도 변화를 확인하기 위해 투여 종료 후 10일, 150일, 300일째에 희생하였다. 투여 종료 후 10일에 희생한 경우 체중에 유의한 차이가 있었지만, 그 외 경우에는 체중과 난소의 무게에 있어서 대조군과 수은 투여군 사이에 유의한 차이가 없었다. 축적된 수은의 위치를 오토메탈로그라피 방법으로 확인하였고, 자궁, 난소, 난자에 축적된 수은 화합물의 위치를 광학현미경으로 분석하였다. 자궁의 경우, 투여 종료 후 10일째에 수은이 지질 세포와 자궁바깥막의 중피에 위치하였다. 150일째에는 수은 농도가 감소하였으며 300일째에는 나타나지 않았다. 투여 종료 후 10일째 난소의 경우, 수은이 피질 부위의 지질 세포와, 난포를 둘러싸는 난포막 세포, 황체에 분포하였다. 150일째에는 수은이 난소의 수질 부위에 축적되었으며, 300일째에는 분포하지 않았다. 투여 종료 후 10과 150일째 난자의 경우, 수은이 난자의 주변부에 주로 분포하였으며, 300일째에는 수은 농도가 감소되고 난자 전체에 고르게 분포하였다. 이런 결과는 암컷 마우스에서 수은에 의해 호르몬 생성, 착상, 그리고 발생 중인 배아가 영향을 받을 수 있다는 사실을 제시해 준다.

• 대한수의학회지
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• 제33권2호
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• pp.337-343
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• 1993

To determine the test conditions to enhance the use of hamster test in dogs, semen were collected from four dogs which had been proven to be fertile in the past and then preserved in BWW (Biggers, Whitten, Whittingham) medium for about 20 hours. The semen were given each different treatment according to the experimental design 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 ova bound with sperm(sperm binding) and ova penetrated by sperm (penetration), and also numbers of both bound and penetrated sperm per ovum. In comparison of different concentrations of canine sperm, the rate of sperm binding was higher in $1.5{\times}10^8$, $1{\times}10^8$, and $5{\times}10^7$ sperm concentrations than $5{\times}10^5$ concentration(p<0.01), and also than $5{\times}10^6$ concentration(p<0.05), respectively. The number of bound sperm per ovum was considerably higher in $1.5{\times}10^8$ sperm concentration than $5{\times}10^7$, $1.5{\times}10^6$, and $5{\times}10^5$ concentrations(p<0.01). The rate of penetration was considerably higher in $1.5{\times}10^8$ and $1{\times}10^8$ sperm concentrations than $5{\times}10^5$ concentration,(p<0.01), and also the higher result of penetration was shown in $5{\times}10^7$ than $5{\times}10^5$ (p<0.05). The number of penetrated sperm per ovum was considerably higher in $1.5{\times}10^8$ sperm concentrations than $5{\times}10^5$(p<0.01), and also the higher number was shown in $1{\times}10^8$ than $5{\times}10^5$ (p<0.05). In comparison of the different preincubation period of canine spermatozoa, no difference was obtained in the results of hamster test among the preincubation periods of 4 hours, 18~24 hours and 48 hours. The canine spermatozoa in BWW medium with $Ca^{2+}$(1.3mM) and without FCS(fetal calf serum), with both $Ca^{2+}$(1.3mM) and FCS, with $Ca^{2+}$(2.6mM) and without FCS, and with both $Ca^{2+}$(2.6mM) and FCS showed no difference in the results of hamster test.These results indicated that the appropriate concentration of sperm should be given in hamster test for dog sperm.

• Clinical and Experimental Reproductive Medicine
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• 제19권2호
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• pp.163-168
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• 1992

This study was carried out to set up the ovum bank for ovum donation and to determine the best freezing method for human immature oocytes. Human immature follicular oocytes were cryopreserved by slow freezing and rapid thawing method. Immature follicular oocytes were treated by propanediol(PROH) solution by 2 and 4 step method in protocols A & B, respectively. In protocol C, immature oocytes were exposed to sucrose prior to treatment of PROH by 4 step method. We compared survival rate, maturation rate, and fertilization rate of immature oocytes among three protocols. Results were as follows. 1. Oocytes treated by the protocol C showed the highest survival rate( 70.3 %) and maturation rate(34.6%) after thawing. 2. Survival rate of oocytes treated by the protocol C was significantly higher than that of the protocol B after thawing(p<0.05). In conclusion, treatment of oocytes with sucrose prior to expose PROH was the best freezing method. Sucrose may have reduced the toxic effect of cryoprotectant to oocytes. We failed to induce fertilization of oocytes, which were treated by any protocols, by conventional insemination method, but obtained 28.8% fertilization rate by using partial zona dissection(PZD) method. This result suggests that micromanipulation(PZD) of the thawed oocytes before insemination will improve the fertilization rate.

• 대한한의학원전학회지
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• 제3권
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• pp.1-150
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• 1989

Oriental medicine thinks life and death as the following. 1. The universe seems to be a kind of organism which is divided into 3 branches, as Heaven, Earth and Man. Man is not created from nihil by the Creator. Heaven and Earth by their interaction operate to produce man. This is similiar that zygote is not created from nihil, and that sperm and ovum are transformed into zygote by their interaction. The symbolic meaning of sperm is Heaven, and that of ovum is Earth. Mind and body, as well as spirit and body, are not the real, but artificial words for the purpose of observing and expressing one man. So there is not spiritual substance as distinct from body. The expected life span of man is subjected to change, and is always becoming through life. Fate, the Creator and the world to come cannot be said to be. 2. After one's death, man is transformend into Heaven and Earth. Dying is this process of transformation. Although man comes into existence and closes one's life, the total life of the universe does not change. The criteria of determination of death is not in cell death, but in somatic death. Somatic death divided into 2 branches, one is heart-lung death, the other is brain death. For the standard of health changes ceaselessly as time goes by, aging and dying is not the process of losing health. Because of mind cannot be seperated from body, we'll feel at ease bodily and mentally in healthy dying. The completion of lifetimes is the value of healthy dying. 3. From the viewpoint of these, we must think to let a person die healthily is the right medical ethics. The way to let a person die healthily is divided into 3 branches, one is treatment, another is prevention and the other is promotion of health. We should treat and prevent death of sickness, but take care of healthy dying.

• 한국동물번식학회:학술대회논문집
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• 한국동물번식학회 2001년도 춘계학술발표대회
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• pp.5-5
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• 2001

This study was to test whether Hanwoo in vitro matured oocytes can be successfully cryopreserved by a new vitrification procedure using MVC method. For the vitrification, oocytes were pretreated in 10% ethylene glycol (EG10) for 5-10 min, exposed in EG30 for 30 sec, each oocytes were individually put on the inner wall of 0.25 $m\ell$ straw, and then straws were directly plunged into L$N_2$. Thawing was taken by 4-step procedures [1.0 Msucrose (MS), 0.5 MS, 0.25 MS, and 0.125 MS] at 37$^{\circ}C$. In vitro developmental capacity (survival, cleavage ($\geq$2-cell) and blastocyst rates) in vitrified group was no significant difference compared to that in other treatment groups (exposed; 100.0, 74.4, 32.3% and control; 100.0, 78.3, 36.3%): high mean percentage of oocytes (91.2%) was survived, 69.4% of them were cleaved and 27.9% of cleaved embryos were developed to blastocyst. Especially, after transfer of in vitro developed embryos in vitrified group, four of six recipient animals were found to pregnant and three of them were ongoing pregnant by manual palpation at 250 days after transfer. However, among them, two healthy female calves (23 and 25kg) were born. This result demonstrates that MVC method is very appropriate freezing method for the Hanwoo in vitro matured oocytes and that ovum bank can be maintained efficiently by MVC cryopreservation method.

• Clinical and Experimental Reproductive Medicine
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• 제44권1호
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• pp.47-51
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• 2017

Objective: This study was designed to report the status of assisted reproductive technology (ART) therapy in South Korea between January 1, 2012 and December 31, 2012. Methods: A localized online survey, originally developed by the International Committee Monitoring Assisted Reproductive Technologies, was first launched and provided to all available ART centers via email in 2015. Fresh embryo transfer (FET) cases were categorized as standard in vitro fertilization, intracytoplasmic sperm injection (ICSI), or half-ICSI. Thawed embryo transfer (TET) and other related procedures, including surgical sperm retrieval, were surveyed. Results: Data from 33,956 ovum pick-up procedures were provided by 75 clinics in 2012. Of the 33,088 cycles in which ovums were retrieved, a complete transfer was performed in 90.5% (29,932 cycles). In addition, 10,079 FET cycles were confirmed to have resulted in clinical pregnancy, representing a pregnancy rate of 30.5% per ovum pick-up and 33.7% per ET. The most common number of embryos transferred in FET was 2 (41.6%), followed by 3 (34.0%), and non-elective single ETs (10.0%). Of the 10,404 TET cycles in which transfer was completed, 3,760 clinical pregnancies (36.1%) were confirmed by ultrasonography. Conclusion: The overall clinical pregnancy rate for FET and TET cycles in 2012 was higher than in 2011 (33.7% vs. 33.2% and 36.1% vs. 31.1%, respectively). The most common number of embryos transferred in FET cycles was 2, unlike in 2011.